Effect of low dose of spermidine on physiological changes in salt-stressed cucumber plants

The present study was aimed to assess the ameliorative potentiality of exogenously applied low dose of spermidine (Spd) (4.0 mL of 0.1 mM) against salt stress in cucumber (Cucumis sativus L.) plants. Salt stress inhibited plant growth, while Spd increased the shoot length and dry weight of leaves in salt-stressed plants. Chlorophyll, carotenoids, and sucrose contents were lower, and the accumulation of superoxide radical was higher in salt-affected plants than in controls, and these detrimental effects were mitigated by Spd treatment. Moreover, salinity diminished the reduced glutathione and total polyphenols and inhibited the activities of catalase, peroxidase, and polyphenol oxidase as compared with controls, and Spd treatment increased all antioxidant activities in salt-injured plants. NaCl-induced oxidative stress caused a significant decrease in GA₄ and GA₅ contents. Spd treatment ameliorated these salt stress effects by increasing the quantities of GA₄. In addition, sodium content was higher and calcium content was lower in salt-treated plants, while Spd treatment reduced the sodium accumulation and increased the calcium level in plants exposed to NaCl. The results suggest that exogenous application of low Spd dose can ameliorate the salt stress effects on cucumber by modulating the components of photosynthetic pigments, antioxidants, gibberellins, and minerals.     

Changes in Oil Accumulation and Fatty Acid Composition of Soybean Seeds under Salt Stress in Response to Salicylic Acid and Jasmonic Acid

greenhouse experiment with factorial arrangement based on randomized complete block design with four replications was conducted in 2015 to evaluate the effects of salicylic acid (SA) (1 mM) and jasmonic acid (JA) (0.5 mM) on oil accumulation and fatty acid composition of soybean oil (Glycine max L.) under salt stress (Non-saline, 4, 7, and 10 dS/m NaCl). Oil percentage of soybean seeds declined, while oil content per seed enhanced with increasing seed filling duration. Foliar application of SA improved oil content per soybean seed at different stages of development under all salinity levels. Although JA treatment enhanced seed oil percentage, oil yield of these plants decreased as a result of reduction in seed yield per plant. In contrast, the highest oil yield was recorded for SA treated plants, due to higher seed yield. Salinity had no significant effects on percentage of palmitic acid and stearic acid, but treatment with JA significantly reduced stearic acid percentage. Oleic acid content of seeds increased, but percentages of linoleic acid, linolenic acid and unsaturation index (UI) of soybean oil decreased with increasing salinity. Foliar application of SA and JA improved oil quality of soybean seeds by reducing oleic acid and enhancing linoleic acid, linolenic acid contents and UI. Exogenous application of SA had the most beneficial effects on soybean seeds due to enhancing oil yield and quality under saline and non-saline conditions.     

Parasitism by Cuscuta pentagona sequentially induces JA and SA defence pathways in tomato

While plant responses to herbivores and pathogens are well characterized, responses to attack by other plants remain largely unexplored. We measured phytohormones and C₁₈ fatty acids in tomato attacked by the parasitic plant Cuscuta pentagona, and used transgenic and mutant plants to explore the roles of the defence‐related phytohormones salicylic acid (SA) and jasmonic acid (JA). Parasite attachment to 10‐day‐old tomato plants elicited few biochemical changes, but a second attachment 10 d later elicited a 60‐fold increase in JA, a 30‐fold increase in SA and a hypersensitive‐like response (HLR). Host age also influenced the response: neither Cuscuta seedlings nor established vines elicited a HLR in 10‐day‐old hosts, but both did in 20‐day‐old hosts. Parasites grew larger on hosts deficient in SA (NahG) or insensitive to JA [jasmonic acid‐insensitive1 (jai1) ], suggesting that both phytohormones mediate effective defences. Moreover, amounts of JA peaked 12 h before SA, indicating that defences may be coordinated via sequential induction of these hormones. Parasitism also induced increases in free linolenic and linoleic acids and abscisic acid. These findings provide the first documentation of plant hormonal signalling induced by a parasitic plant and show that tomato responses to C. pentagona display characteristics similar to both herbivore‐ and pathogen‐induced responses.     

Silicon: a duo synergy for regulating crop growth and hormonal signaling under abiotic stress conditions

Abiotic stresses, such as salinity, heavy metals and drought, are some of the most devastating factors hindering sustainable crop production today. Plants use their own defensive strategies to cope with the adverse effects of these stresses, via the regulation of the expression of essential phytohormones, such as gibberellins (GA), salicylic acid (SA), jasmonates (JA), abscisic acid (ABA) and ethylene (ET). However, the efficacy of the endogenous defensive arsenals of plants often falls short if the stress persists over an extended period. Various strategies are developed to improve stress tolerance in plants. For example, silicon (Si) is widely considered to possess significant potential as a substance which ameliorate the negative effects of abiotic stresses, and improves plant growth and biomass accumulation. This review aims to explain how Si application influences the signaling of the endogenous hormones GA, SA, ABA, JA and ET during salinity, wounding, drought and metal stresses in crop plants. Phytohormonal cross talk plays an important role in the regulation of induced defences against stress. However, detailed molecular and proteomic research into these interactions is needed in order to identify the underlying mechanisms of stress tolerance that is imparted by Si application and uptake.     

Silencing genes encoding omega-3 fatty acid desaturase alters seed size and accumulation of Bean pod mottle virus in soybean

Omega-3 fatty acid desaturase (FAD3)-catalyzed conversion of linoleic acid to linolenic acid (18:3) is an important step for the biosynthesis of fatty acids as well as the phytohormone jasmonic acid (JA) in plants. We report that silencing three microsomal isoforms of GmFAD3 enhanced the accumulation of Bean pod mottle virus (BPMV) in soybean. The GmFAD3-silenced plants also accumulated higher levels of JA, even though they contained slightly reduced levels of 18:3. Consequently, the GmFAD3-silenced plants expressed JA-responsive pathogenesis-related genes constitutively and exhibited enhanced susceptibility to virulent Pseudomonas syringae. Increased accumulation of BPMV in GmFAD3-silenced plants was likely associated with their JA levels, because exogenous JA application also increased BPMV accumulation. The JA-derived increase in BPMV levels was likely not due to repression of salicylic acid (SA)-derived signaling because the GmFAD3-silenced plants were enhanced in SA-dependent defenses. Furthermore, neither exogenous SA application nor silencing the SA-synthesizing phenylalanine ammonia lyase gene altered BPMV levels in soybean. In addition to the altered defense responses, the GmFAD3-silenced plants also produced significantly larger and heavier seed. Our results indicate that loss of GmFAD3 enhances JA accumulation and, thereby, susceptibility to BPMV in soybean.     

Nitric oxide enhances salt tolerance in cucumber seedlings by regulating free polyamine content

Nitric oxide (NO), an endogenous signaling molecule in plants and animals, mediates responses to abiotic and biotic stresses. This study was conducted in nutrient solution to investigate the effects of exogenous sodium nitroprusside (SNP), an NO donor, on plant growth and free polyamine content in cucumber leaves and roots under NaCl stress. The results showed that 100 μM SNP in solution significantly improved the growth of cucumber seedlings under NaCl stress for 8 days, as indicated by increased, plant height, stem thickness, fresh weight and increased dry matter accumulation. Further analysis demonstrated that the content of free polyamines and the activity of polyamine oxidase (PAO) in cucumber seedling leaves and roots initially increased dramatically under NaCl stress, although they decreased over a longer period of stress. Throughout the treatment period, the value of (spermine + spermidine)/putrescine [(Spd + Spm)/Put] also decreased under NaCl stress compared to the control. In contrast, the application of 100 μM SNP in the nutrient solution decreased the content of free Put, Spd, total free polyamines and PAO activity under NaCl stress. It also caused an increase in the content of Spm and the value of (Spd + Spm)/Put, adjusted the ratio of three kinds of free polyamines (Put, Spd, Spm) in cucumber seedlings. The high (Spd + Spm)/Put value and the accumulation of Spm were beneficial to improving the salt tolerance of plants. Therefore, NO alleviated the damage to cucumber seedlings caused by salt stress. NO enhanced the tolerance of cucumber seedlings to NaCl stress by regulating the content and proportions of the different types of free polyamines.     

Simultaneous quantitative LC-ESI-MS/MS analyses of salicylic acid and jasmonic acid in crude extracts of Cucumis sativus under biotic stress

Salicylic acid (SA) and jasmonic acid (JA) are plant hormones involved in basal resistance against plant pathogens and also in induced resistance. The aim of this study is to develop a fast and sensitive method to determine simultaneously the levels of both these hormones. The present paper proposes a method that includes hormone extraction with MeOH-H(2)O-HOAc (90:9:1, v/v), evaporation of the extracts, and injection into the liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) system in multiple reaction monitoring (MRM). Endogenous SA and JA levels in noninfested control cucumber cotyledons were 30.96 and 0.73ngg(-1) fresh weight, respectively. In roots, the levels were 8.31 and 15.82ngg(-1) FW, respectively. In plants treated with the biological control agent Trichoderma asperellum strain T-34, the levels of SA and JA did not differ from control plants. Rhizoctonia solani-diseased cucumber plants showed higher levels of SA and JA compared to noninfested controls (up to 2 and 13-fold higher, respectively). Detection limits for SA and JA were 0.45 and 0.47ngg(-1) fresh weight, respectively. The results of our research include the development of a method that is both fast and highly sensitive in the simultaneous quantitation of SA and JA from crude cucumber plant extracts, avoiding any purification and derivatization steps.     

Different relative humidity conditions combined with chloride and sulfate salinity treatments modify abscisic acid and salicylic acid levels in the halophyte Prosopis strombulifera

It has been shown that abscisic acid (ABA) and salicylic acid (SA) act as endogenous signal molecules responsible for inducing abiotic stress tolerance in plants. However, our knowledge on the role of both phytohormones in response to environmental conditions in halophytic plants is still limited. In this study endogenous ABA and SA levels, growth parameters and chlorophylls content were determined in leaves and roots of the halophyte Prosopis strombulifera cultivated under increasing NaCl and Na₂SO₄ concentrations, at 30 and 70 % relative humidity (RH) conditions. Endogenous ABA and SA content differed depending on the salt type and concentration, RH, plant age and the organ analyzed. Under low RH conditions P. strombulifera growth was strongly inhibited and chlorophyll a and b content were decreased. In leaves of Na₂SO₄-treated plants at 30 % RH, high ABA levels were correlated with protection against dehydration and ion toxicity. Instead, high SA levels were correlated with the damaging effect of sulfate anion and low RH on plant growth. NaCl-treated plants growth was also inhibited at 30 % RH although levels of both hormones were not significantly increased. Taken together, the salt toxic effects on growth parameters and photosynthetic pigments were accentuated by low RH conditions and these responses were reflected on ABA and SA content.     

Effects of 5-aminolevulinic acid on nitrogen metabolism and ion distribution of watermelon seedlings under salt stress

The effects of foliar spray application of 5-aminolevulinic acid (ALA) on the growth, nitrogen metabolism, and ion distribution of salt-stressed watermelon (Citrullus lanatus (Thunb.) Matsum. and Nakai) seedlings were investigated. Supplementation of the nutrient solution with 100 mM NaCl significantly reduced leaf and root biomass of watermelon plants. Foliar application of 1.25 mM ALA significantly alleviated the inhibition of plant growth under salt stress. Salinity induced significant accumulation of nitrate, ammonium, and soluble protein and a significant decrease in the activities of nitrate reductase (NR), nitrite reductase (NiR), glutamine synthetase (GS), glutamate synthase (GOGAT), and glutamate dehydrogenase (GDH) in watermelon plants. However, ALA significantly increased the activities of NR, GS, GOGAT, and GDH, but decreased the ammonium content and NiR activity. In addition, salt stress resulted in significant accumulation of Na⁺ and Cl^? in plants, but decreased the contents of K⁺ and Mg²⁺. Application of ALA alleviated the salt stress-induced ion toxicity, and increased the contents of K⁺ and Mg²⁺. ALA also increased soluble protein and proline contents in salt-stressed watermelon plants. These results indicated that application of ALA alleviated the accumulation of Na⁺ and Cl^? in salt-stressed watermelon plants, especially through regulating nitrogen metabolism and ion distribution, which were associated with an improvement in plant growth.     

Herbivore-induced rice resistance against rice blast mediated by salicylic acid

In agro-ecosystems,plants are important mediators of interactions between their associated herbivorous insects and microbes,and any change in plants induced by one species may lead to cascading effects on interactions with other species.Often,such effects are regulated by phytohormones such as jasmonic acid(JA)and salicylic acid(SA).Here,we investigated the tripartite interactions among rice plants,three insect herbivores(Chilo suppressalis,Cnaphalocrocis medinalis or Nilapai-vata lugens),and the causal agent of rice blast disease,the fungus Magnaporthe oryzae.We found that pre-infestation of rice by C.suppressalis or N.lugens but not by C.medinalis conferred resistance to M.oryzae.For C.suppressalis and N.lugens,insect infestation without fungal inoculation induced the accumulation of both JA and SA in rice leaves.In contrast,infestation by C.medinalis increased JA levels but reduced SA levels.The exogenous application of SA but not of JA conferred resistance against M.oryzae.These results suggest that preinfestation by C suppressalis or N.lugens conferred resistance against M.oryzae by increasing SA accumulation.These findings enhance our understanding of the interactions among rice plant,insects and pathogens,and provide valuable information for developing an ecologically sound strategy for controlling rice blast.     

Changes in phytohormones and fatty acids in wheat and rice seedlings in response to Hessian fly (Diptera: Cecidomyiidae) infestation

Phytohormones and fatty acids (FAs) play important roles in plant resistance to insects and pathogens. In this study, we investigated the similarities and differences in the accumulations of phytohormones and FAs in the resistant wheat (Triticum aestivum L.) 'Molly' and the nonhost rice (Oryza sativa L.) 'Niponbare' in responses to Hessian fly, Mayetiola destructor (Say) (Diptera: Cecidomyiidae), larval attacks. Using chemical ionization-gas-chromatography/mass spectrometry, we analyzed the concentrations of 13 phytohomones and FAs at the attack site of wheat and rice plants at 1, 6, 24, or 48 h after the initial attack. Hessian fly attack resulted in increases of salicylic acid (SA), 12-oxo-phytodienoic acid (OPDA), palmitic acid (FA16:0), but a decrease of abscisic acid in both wheat and rice plants. In addition, the accumulation of jasmonic acid (JA) increased, whereas the accumulation of cinnamic acid (CA) decreased in wheat plants, but no changes were observed in the accumulation of JA, and the accumulation of CA increased in rice plants after Hessian fly attack. However, the accumulations of benzoic acid, strearic acid (FA18:0), and oleic acid (FA18:1) increased in rice plants, but no changes were observed in wheat plants after Hessian fly attack. Hessian fly-induced changes were more rapid in wheat plants in comparison with those in rice plants. Our study suggests that SA and OPDA may be involved in resistance of wheat and rice plants to Hessian fly and that the R gene-mediated resistance responses are more rapid than nonhost resistance responses.     

New perspective on the mechanism of alleviating salt stress by spermidine in barley seedlings

Salt stress is considered to be a major limiting factor for plant growth and crop productivity. Salt injuries in plants are mostly due to excess Na⁺ entry. A possible survival strategy of plants under saline environments is the effective compartmentation of excess Na⁺ by sequestering Na⁺ in roots and inhibiting transport of Na⁺ from roots to shoots. Our previous study showed that exogenous application of polyamines (PAs) could attenuate salt injuries in barley plants. In order to further understand such protective roles of PAs against salt stress, the effects of spermidine (Spd) on sodium and potassium distribution in barley (Hordeum vulgare L.) seedlings under saline conditions were investigated. The results showed that exogenous application of Spd induced reductions in Na⁺ levels in roots and shoots with comparison of NaCl-treated plants, while no significant changes in K⁺ levels were observed. Correspondingly, the plants treated with Spd exogenously maintained high values of [K⁺]/[Na⁺] as compared with salt-stressed plants. Moreover, it was shown by X-ray microanalysis that K⁺ and Na⁺ accumulated mainly in the exodermal intercellular space and cortical cells of roots under salinity stress, and low accumulation was observed in endodermal cells and stelar parenchyma, indicating Casparian bands possibly act as ion transport barriers. Most importantly, Spd treatment further strengthened this barrier effects, leading to inhibition of Na⁺ transport into shoots. These results suggest that, by reinforcing barrier effects of Casparian bands, exogenous Spd inhibits Na⁺ transport from roots to shoots under conditions of high salinity which are beneficial for attenuating salt injuries in barley seedlings.     

Spermidine and related-metabolic inhibitors modulate sugar and amino acid levels in Vitis vinifera L.: possible relationships with initial fruitlet abscission

The relationships between free polyamines (PAs), sugar and amino acid status were investigated in cuttings from two cultivars of Vitis vinifera L., Pinot noir (PN), a low abscising cultivar and Merlot (MRT), a high abscising one. In both cultivars free PAs decreased in inflorescences, but more drastically in MRT plants. Upon anthesis, this was associated with a decreased sugar content, especially sucrose, and an increase in total free amino acids. Thereafter, sucrose and amino acids showed opposite trends. In addition, darkening the PN plants at full flowering resulted in a dramatic decrease of PAs and sugars in inflorescences, but an increase in amino acid content, followed by high abscission. The concept that initial fruitlet abscission might be regulated by free PAs through changes in primary metabolites was hypothesized. Hence, the application of exogenous spermidine (Spd), but not putrescine (Put), prior to flowering markedly inhibits abscission. The Spd treatment also increased soluble sugar content but reduced amino acids in both leaves and inflorescences, while Put had no significant effect. By contrast, cyclohexylamine and beta-hydroxyethylhydrazine, as potent inhibitors of Spd synthase and PA-oxidases, respectively, exerted inverse effects on sugar, amino acid and abscission levels. Sucrose and free proline seemed to be highly sensitive to these treatments. This study suggests that Spd could regulate fruitlet abscission in grapevine by modulating, in a reverse way, the levels of sugars and amino acids in inflorescences.     

Adaptive mechanism of Echinochloa crus-galli Beauv. var. formosensis Ohwi under salt stress: Effect of salicylic acid on salt sensitivity

Our previous study revealed that salicylic acid (SA) accumulates in salt-stressed rice (Oryza sativa L. cv. Nipponbare) seedlings, and we hypothesized that the accumulation of SA might potentiate oxidative injury in rice seedlings since the inhibition of SA synthesis alleviated the growth inhibition under high salinity. To further clarify the action of SA under salt stress, we investigated the changes in the SA content, the activities of the antioxidative enzymes, and the effects of exogenous SA on barnyardgrass (Echinochloa crus-galli Beauv. var. formosensis Ohwi), a gramineous weed which shows lower SA content and is more salt tolerant than rice. In E. crus-galli seedlings exposed to high salinity, neither free nor conjugated SA content showed any increase, while the fresh weight of the shoot and chlorophyll fluorescence (Φ_PSII) slightly decreased. When E. crus-galli seedlings were treated with salt after foliar application of SA, the absorbed SA resulted in the enhancement of the salt-induced growth inhibition and a striking reduction of the Φ_PSII value. Catalase (CAT) and superoxide dismutase (SOD) activities of E. crus-galli seedlings were induced by the salt treatment. However, SA pre-treatment suppressed such an induction of CAT activity and further promoted SOD activity, both of which led to the elevation of the leaf hydrogen peroxide (H₂O₂) level. The present results suggested that enlargement of the cellular SA pool facilitates the generation of H₂O₂ through the suppression of CAT activity and through a remarkable promotion of SOD activity, and thereby enhances the oxidative injury caused by salt stress.     

Ozone-induced expression of the Arabidopsis FAD7 gene requires salicylic acid, but not NPR1 and SID2

The Arabidopsis FAD7 gene encodes a plastid omega-3 fatty acid desaturase that catalyzes the desaturation of dienoic fatty acids to trienoic fatty acids in chloroplast membrane lipids. The expression of FAD7 was rapidly and locally induced by ozone exposure, which causes oxidative responses equivalent to pathogen-induced hypersensitive responses and subsequently activates various defense-related genes. This induction was reduced in salicylic acid (SA)-deficient NahG plants expressing SA hydroxylase, but was unaffected in etr1 and jar1 mutants, which are insensitive to ethylene and jasmonic acid (JA), respectively. The SA dependence of the FAD7 induction was confirmed by the exogenous application of SA. SA-induced expression of FAD7 in the npr1 mutant which is defective in an SA signaling pathway occurred to the same extent as in the wild type. Furthermore, in the sid2 mutant which lacks an enzyme required for SA biosynthesis, the expression of FAD7 was induced by ozone exposure. These results suggest that the ozone-induced expression of FAD7 gene requires SA, but not ethylene, JA, NPR1 and SID2.     

Impact of salicylic acid on the antioxidant enzyme system and hydrogen peroxide production in Cucumis sativus under chilling stress

Salicylic acid (SA) is a naturally produced compound and has been implicated to play important roles in defense of plants against diverse biotic and abiotic stresses. To understand how SA functions in the tolerance of cucumber (Cucumis sativus) to chilling stress, endogenous SA levels in two different cultivars with opposite chilling responsiveness were quantified. Membrane integrity, including malondialdehyde (MDA) content and leakage of electrolyte, was also examined in SA-pretreated cucumber plants under chilling conditions. In addition, activities of the two antioxidant enzymes peroxidase (POD) and catalase (CAT) were quantified, and hydrogen peroxide (H2O2) production was investigated histochemically in SA-treated leaves under chilling temperature. Chilling stress resulted in greater induction of SA levels in the chilling-tolerant cultivar Changchun mici in both leaves and seeds compared to the chilling-sensitive one Beijing jietou, while the former one contained higher levels of SA than the latter one in the seeds under normal conditions. Pretreatment with SA diminished the increased electrolyte leakage and MDA content caused by chilling in the leaves of both cultivars, while much less MDA and electrolyte leakage were produced in Changchun mici compared to Beijing jietou. Moreover, exogenous application of SA increased significantly the POD and CAT activities and soluble protein content. Most importantly, exogenous SA treatment could eliminate the accumulation of H202 in leaves and cotyledons of both cultivars caused by chilling stress. The data clearly demonstrated that the chilling-tolerant cultivar displays a higher SA level than the chilling-sensitive one, and that exogenous SA can enhance the chilling tolerance ability, which might be achieved through modulating the antioxidant system in cucumber.