Amelioration of salinity stress by exogenously applied spermidine or spermine in three varieties of indica rice differing in their level of salt tolerance

We present here the comparative protective potentiality of exogenously applied polyamines (PAs), namely spermidine (Spd) and spermine (Spm), in mitigating NaCl toxicity and inducing short-term salinity tolerance in three indica rice varieties, namely M-1-48 (salt-sensitive), Nonabokra (salt-tolerant) and Gobindobhog (highly sensitive). The retardation in root length or shoot length and toxic Na⁺ accumulation or K⁺ loss, the considerable increment in malondialdehyde/H₂O₂ accumulation or lipoxygenase activity, all of which were particularly noteworthy in M-1-48 and Gobindobhog during salinity stress, was appreciably reduced by co-treatment with Spd or Spm. Both the PAs also inhibited the extent of salt-induced protein carbonylation in all the varieties and enhanced protease activity, especially in Gobindobhog. The prevention of chlorophyll degradation was better with Spd in Nonabokra and Gobindobhog. While the salt-induced increase in anthocyanin or reducing sugar level was further prompted by Spd or Spm in all the varieties, the proline content was elevated by Spd particularly in Gobindobhog. During salinity stress, both the PAs were effective in lowering the putrescine accumulation in M-1-48 and Gobindobhog, and strikingly increasing the Spm level in all the varieties, the highest being in Gobindobhog. In addition, they enhanced the activity of peroxidases and compensated for the decreased catalase activity in all the varieties. Thus the two PAs could recuperate all the three varieties from salt-induced damages to different degrees. The salt injuries, encountered in M-1-48 and Gobindobhog, both of which showed greater susceptibility to salinity stress, were more pronouncedly alleviated and counteracted by the PAs, than the salt-tolerant Nonabokra. The reversal of inhibitory effect of salinity stress was conferred by preventing growth inhibition or various forms of cellular damages, maintaining proper K⁺/Na⁺ balance or triggering the level of osmolytes and activity of antioxidant enzymes. Our communication offers a referenced evidence for an understanding of the mechanism by which higher PAs relieve the damages particularly in salt-sensitive rice varieties.     

Exogenous Putrescine Reduces Sodium and Chloride Accumulation in NaCl-Treated Calli of the Salt-Sensitive Rice Cultivar I Kong Pao

In order to gain information on the putative involvement of polyamines (PAs) in the response of rice cells to salinity, mature embryo-derived calli issued from the salt-sensitive cultivar I Kong Pao were exposed for 3 months to the simultaneous presence of NaCl (0, 150 and 300 mM) and exogenous polyamines (putrescine (Put): 1 and 10 mM; spermidine (Spd): 1 and 10 mM; spermine (Spm): 1 mM). Callus growth, endogenous PAs, Na⁺, K⁺ and Cl⁻ concentrations were quantified and analysed in relation to cell viability based on 2,3,5-triphenytetrazolium chloride (TTC) reduction. All exogenous PAs were efficiently absorbed from the external medium. Exogenous Put 1 mM clearly stimulated growth of salt-stressed calli in relation to a decrease in both Na⁺ and Cl⁻ accumulation. In contrast, Spd 10 mM and Spm 1 mM exacerbated the deleterious impact of NaCl on callus growth and induced a decrease in K⁺ concentration. While Put helped in the maintenance of cell viability, Spd 10 mM and Spm 1 mM decreased cell viability, mainly in relation to an inhibition of the alternative respiratory pathway. It is proposed that Put may assume positive functions in salt stress resistance in rice.     

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.     

Involvement of polyamines, abscisic acid and anti-oxidative enzymes in adaptation of Blue Panicgrass (Panicum antidotale Retz.) to saline environments

A hydroponic experiment was conducted to assess the possible involvement of polyamines (PAs), abscisic acid (ABA) and anti-oxidative enzymes such as superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) in adaptation of six populations of Panicum antidotale Retz. to selection pressure (soil salinity) of a wide range of habitats. Plants of six populations were collected from six different habitats with ECe ranging from 3.39 to 19.23 dS m⁻¹ and pH from 7.65 to 5.86. Young tillers from 6-month-old plants were transplanted in plastic containers each containing 10 l of half strength Hoagland's nutrient solution alone or with 150 mol m⁻³ NaCl. After 42 days growth, contents of polyamines (Put, Spd and Spm) and ABA, and the activities of anti-oxidative enzymes (SOD, POD and CAT) of all populations generally increased under salt stress. The populations collected from highly saline habitats showed a greater accumulation of polyamines and ABA and the activities of anti-oxidative enzymes as compared to those from mild or non-saline habitats. Moreover, Spm/Spd and Put/(Spd + Spm) ratios generally increased under salt stress. However, the populations from highly saline environments had significantly higher Spm/Spd and Put/(Spd + Spm) ratios as compared to those from mild or non-saline environments. Similarly, the populations adapted to high salinity accumulated less Na⁺ and Cl⁻ in culm and leaves, and showed less decrease in leaf K⁺ and Ca²⁺ under salinity stress. Higher activities of anti-oxidative enzymes and accumulation of polyamines and ABA, and increased Spm/Spd and Put/(Spm + Spd) ratios were found to be highly correlated with the degree of adaptability of Panicum to saline environment.     

Spermine Pre-Treatment Improves Some Physiochemical Parameters and Sodium Transporter Gene Expression of Pumpkin Seedlings under Salt Stress

It is known that polyamines (PAs) including spermine (Spm) enhance the abiotic stress tolerance of crops. Here, the effects of hydroponic Spm pre-treatment on the amelioration of the adverse effects of salinity were investigated in pumpkin (Cucurbita pepo L.) that is an economically important horticultural crop and sensitive to salinity, especially at the establishment stage. For this purpose, 10-day-old, uniform-sized seedlings were transplanted to plastic containers containing Hoagland nutrient solution. Spm was added at 0.1 and 1 mM to the hydroponic medium for 5 days before stress. The plants were treated with 40 and 80 mM NaCl for inducing salinity stress. Salt stress reduced the plant growth and potassium content in roots, and these detrimental effects were alleviated when plants were pre-treated with Spm. Salinity stress caused a significant increase in sodium and γ-amino butyric acid (GABA) content when compared with controls. Spm pre-treatment ameliorated these salinity stress effects by increasing sodium content of root and leaves GABA content. Expression analysis of two sodium transporter genes, salt overly sensitive1 (SOS1) and Na⁺/H⁺ exchanger (NHX1) revealed that their expression was differentially induced in roots of plants treated either with salinity or Spm. These results suggest that Spm via the overexpression of the NHX1 gene substantially increased the tolerance to stress by sequestering excess Na+ into the vacuoles and sustaining a better cellular environment. Moreover, Spm has potential to scavenge directly free radical and to alleviate growth inhibition and promote the activity of antioxidant system enzymes in pumpkin seedlings under salt stress.     

Effect of arginine and urea on polyamines content and growth of bean under salinity stress

Prior to sowing, seeds of bean (Phaseolus vulgaris L.) were treated with 4 mM arginine or 0.1% urea, as nitrogen source. The seeds were then subjected to salinity stress. Arginine and urea treatments stimulated germination of both unstressed and salinity-stressed seeds. It was interesting to observe that the increased germination percentage in response to arginine and urea treatments was associated with increased content of polyamines, particularly putrescine (Put), spermidine (Spd) and spermine (Spm). Growth of the seedlings was also improved by application of arginine and urea, which was also associated with increased content of the polyamines Spd and Spm, while the Put content decreased. Total soluble sugars were much accumulated in response to arginine and urea treatments under salinity stress for cellular osmoregulation. The ratio of K⁺/Na⁺ increased in the leaves by application of arginine and urea, indicating a more alleviation to the adverse effects of salinity stress. Changes in proteinogenic amino acids were also investigated.     

Involvement of polyamines,abscisic acid and anti-oxidative enzymes in adaptation of Blue Panicgrass (Panicum antidotale Retz.) to saline environments

A hydroponic experiment was conducted to assess the possible involvement of polyamines (PAs), abscisic acid (ABA) and anti-oxidative enzymes such as superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) in adaptation of six populations of Panicum antidotale Retz. to selection pressure (soil salinity) of a wide range of habitats. Plants of six populations were collected from six different habitats with ECe ranging from 3.39 to 19.23 dS m⁻¹ and pH from 7.65 to 5.86. Young tillers from 6-month-old plants were transplanted in plastic containers each containing 10 l of half strength Hoagland's nutrient solution alone or with 150 mol m⁻³ NaCl. After 42 days growth, contents of polyamines (Put, Spd and Spm) and ABA, and the activities of anti-oxidative enzymes (SOD, POD and CAT) of all populations generally increased under salt stress. The populations collected from highly saline habitats showed a greater accumulation of polyamines and ABA and the activities of anti-oxidative enzymes as compared to those from mild or non-saline habitats. Moreover, Spm/Spd and Put/(Spd + Spm) ratios generally increased under salt stress. However, the populations from highly saline environments had significantly higher Spm/Spd and Put/(Spd + Spm) ratios as compared to those from mild or non-saline environments. Similarly, the populations adapted to high salinity accumulated less Na⁺ and Cl⁻ in culm and leaves, and showed less decrease in leaf K⁺ and Ca²⁺ under salinity stress. Higher activities of anti-oxidative enzymes and accumulation of polyamines and ABA, and increased Spm/Spd and Put/(Spm + Spd) ratios were found to be highly correlated with the degree of adaptability of Panicum to saline environment.     

Effects of exogenous polyamines on nitrate tolerance in cucumber

Putrescine (Put), spermidine (Spd), and spermine (Spm) are the major polyamines (PAs) in plant, which are not only involved in the regulation of plant developmental and physiological processes, but also play key roles in modulating the defense response of plants to diverse environmental stresses. In this study, Cucumis sativus L. seedlings were cultivated in nutrient solution and sprayed with three kinds of PAs (Put, Spd, and Spm). The effects of PAs were investigated on excess nitrate stress tolerance of C. sativus by measuring growth and nitrogen (N) metabolism parameters. The contents of NO_3-^?N, NH_4-⁺N, proline and soluble protein in leaves were increased; while plant height, leaf area, shoot fresh and dry weight, root fresh weight were decreased under 140 mM NO₃^? treatment for 7 d. In addition, the activities of nitrate reductase (NR), glutamate synthase (GOGAT), and glutamate dehydrogenase (GDH) were significantly inhibited under 140 mM NO₃^? treatment for 7 d. With foliar treatment by 1 mM Spd or Spm under stress treatment, the contents of Spm, Put, and Spd in leaves increased significantly, except that Spm content decreased under Spd treatment. The activities of NR, glutamine synthetase (GS), GOGAT and GDH and plant height, leaf area, shoot fresh and dry weights were significantly increased. The contents of proline and soluble protein in leaves were significantly enhanced. In contrast, the accumulation of NO_3-^?N and NH_4-⁺N were significantly decreased. However, there were minor differences in activities of N metabolism enzymes and the content of osmotic adjustment substances under 1 mM Put treatment. These findings suggest that 1 mM exogenous Spm or Spd could enhance the capacity of N metabolism, promote growth and increase resistance to high concentrations of NO₃^?. The ameliorating effect of Spd was the best, and that of Put the worst.     

Protective role of exogenous polyamines on salinity-stressed rice (Oryza sativa) plants

Salt-tolerant Pokkali rice plants accumulate higher polyamines (PAs) such as spermidine (Spd) and spermine (Spm) in response to salinity stress, while the sensitive cultivar M -1–48 is unable to maintain high titres of these PAs under similar conditions. The effects of the triamine Spd and the tetramine Spm on physiological and biochemical changes in 12-day-old rice seedlings were investigated during salinity stress to determine whether they could protect the sensitive plants from stress effects. At physiological concentrations Spd and Spm significantly prevented the leakage of electrolytes and amino acids from roots and shoots induced by salinity stress. To different degrees they also prevented chlorophyll loss, inhibition of photochemical reactions of photosynthesis as well as downregulation of chloroplast-encoded genes like psbA , psbB , psbE and rbcL , indicating a positive correlation between salt tolerance and accumulation of higher PAs in rice. The inhibitory effect of salinity stress and its reversal by exogenous PAs were more pronounced in the salt-sensitive M -1–48 plants than in the tolerant Pokkali plants.     

Protective effect of exogenous polyamines on root tonoplast function against salt stress in barley seedlings

Salinity stress is one of the most serious factors limiting the productivity of agricultural crops. A possible survival strategy of plants under saline conditions is to sequester excess Na⁺ in the vacuole by vacuolar Na⁺/H⁺ antiport using a pH gradient generated by H⁺-ATPasc (EC 3.6.1.35) and H⁺-Pyrophosphatase (H⁺-PPase; EC 3.6.1.1) to maintain a higher K⁺/Na⁺ ratio in cytoplasm. The effect of exogenously applied polyamines (PAs) in stabilizing root tonoplast integrity and function against salt stress in the barley (Hordeum vulgare L.) seedlings was investigated. The NaCl-induced reductions in the contents of phospholipids and PAs in tonoplast vesicles isolated from barely seedling roots, as well as the activities of H⁺-ATPase, H⁺-PPase and vacuolar Na⁺/H⁺ antiport were all partially restored by the application of 0.5 mM putrescine and 0.5 mM spermidine, especially the former. The above results indicated that one of the mechanisms involved in attenuating salt injury in barley seedlings by exogenous PAs application was to maintain tonoplast integrity and function under saline conditions. Moreover, the possible mechanism involved in counteracting detrimental effects of salt on the barley seedlings by the application of exogenous PAs was discussed.     

Response of Cleopatra mandarin seedlings to a polyamine-biosynthesis inhibitor under salt stress

Six-month-old, uniform-sized seedlings of Cleopatra mandarin growing in hydroponics in Hoagland nutrient solution under glasshouse conditions were subjected to salinity treatment (NaCl 75 mM). Addition of NaCl to the nutrient medium reduced plant growth (shoot height and leaf number), leaf chlorophyll content, chlorophyll fluorescence yields (Fv/Fm), net photosynthesis, stomatal conductance, transpiration rate, intracellular CO₂ concentration, N, K⁺ and Ca²⁺ + Mg²⁺ contents of the leaves; and increased total putrescine (Put), proline and Na⁺ and Cl⁻ contents of the leaves. Addition of d-arginine (1 mM) to the saline medium further reduced the values of all the parameters except Fv/Fm, photosynthesis and related parameters and Ca²⁺ + Mg²⁺ contents of the leaves, and increased total spermine (Spm) content of the leaves. However, total Put contents were decreased; spermidine (Spd) contents and Na⁺ and Cl⁻ titres of the leaves were unaffected. Weekly spray of d-arginine (5 mM) was less harmful than its addition to the nutrient medium (1 mM). Addition of 0.5 mM Spd to the medium alleviated the adverse effects of d-arginine by improving plant growth, leaf chlorophyll content, total Put, Spd, Spm, N, P, K⁺ and Ca²⁺ + Mg²⁺ contents of the leaves. Weekly spray of Spd (5 mM) was less effective than its addition in the nutrient medium (0.5 mM).     

Spermidine alleviates the growth of saline-stressed ginseng seedlings through antioxidative defense system

Protective effects of exogenous spermidine (Spd), activity of antioxygenic enzymes, and levels of free radicals in a well-known medicinal plant, Panax ginseng was examined. Seedlings grown in salinized nutrient solution (150 mM NaCl) for 7 d exhibited reduced relative water content, plant growth, increased free radicals, and showing elevated lipid peroxidation. Application of Spd (0.01, 0.1, and 1 mM) to the salinized nutrient solution showed increased plant growth by preventing chlorophyll degradation and increasing PA levels, as well as antioxidant enzymes such as CAT, APX, and GPX activity in the seedlings of ginseng. During salinity stress, Spd was effective for lowering the accumulation of putrescine (Put), with a significant increase in the spermidine (Spd) and spermine (Spm) levels in the ginseng seedlings. A decline in the Put level ran parallel to the higher accumulation of proline (Pro), and exogenous Spd also resulted in the alleviation of Pro content under salinity. Hydrogen peroxide (H₂O₂) and superoxide (O₂⁻) production rates were also reduced in stressed plants after Spd treatment. Furthermore, the combined effect of Spd and salt led to a significant increase in diamine oxidase (DAO), and subsequent decline in polyamine oxidase (PAO). These positive effects were observed in 0.1 and 1 mM Spd concentrations, but a lower concentration (0.01 mM) had a very limited effect. In summary, application of exogenous Spd could enhance salt tolerance of P. ginseng by enhancing the activities of enzyme scavenging system, which influence the intensity of oxidative stress.     

Soil Salinity and Water Stress and Their Effect on Susceptibility to Verticillium Wilt Disease, Ion Composition and Growth of Pistachio

The effects of soil salinity and water stress on Verticillium wilt, ion composition and growth of pistachio were studied in a greenhouse experiment (18–32°C). Treatments consisted of three levels of salinity (0, 1200 and 2400 mg NaCl/kg soil), three levels of water stress (3, 7 and 14 day irrigation regimes) and two Pistachio cultivars (Sarakhs and Qazvini, common rootstocks in Iran). Infested soil containing 50 microsclerotia/g of a pistachio isolate of Verticillium dahliae was used for all treatments and non‐infested soils were used as control. The experiment was arranged in a completely randomized design with three replications. Eight‐week‐old pistachio seedlings were transferred to infested and non‐infested soil and then exposed to salt stress and thereafter water stress. Shoot dry weights of both rootstocks were reduced significantly with increasing NaCl levels; however, increasing irrigation regimes reduced salt injury. Salt stress significantly increased shoot and root colonization by V. dahliae in both cultivars. Moreover, increasing of salinity level was positively correlated with increasing concentrations of Na⁺, K⁺ and Cl^? in both cultivars, but negatively correlated with increasing irrigation regimes. Based on these results, Sarakhs and Qazvini were found to be sensitive and tolerant to the effect of irrigation regimes, salinity and Verticillium wilt disease, respectively. Although there were no interactive effects of irrigation and salinity on V. dahliae infection.     

Exogenous spermidine and spermine enhance cadmium tolerance of <Emphasis Type="Italic">Potamogeton malaianus</Emphasis>

In order to investigate the effects of spermidine (Spd) and spermine (Spm) on cadmium stress, the content of chlorophyll, hydrogen peroxide (H₂O₂), malondialdehyde (MDA), soluble protein and proline, the rate of O₂^·− generation, and activities of antioxidant enzymes (superoxide dismutase (SOD), peroxidase (POD), ascorbate peroxidase (APX), and glutathione reductase (GR)) in Potamogeton malaianus Miq. were measured. Exogenous application of Spd or Spm significantly enhanced the level of proline, retarded the loss of chlorophyll, enhanced photosynthesis, decreased the rate of O₂^·− generation and H₂O₂ content, and prevented Cd-induced lipid peroxidation. Spd and Spm also effectively maintained the balance of antioxidant enzyme activities under Cd stress; however, GR activity was found to increase only slightly in response to polyamines (PAs). The antioxidant systems, which were modified by PAs, were able to moderate the radical-scavenging system and to lessen in this way the oxidative stress. These results suggest that both Spd and Spm can enhance Cd tolerance of P. malaianus.     

Protective Action of Spermine and Spermidine against Photoinhibition of Photosystem I in Isolated Thylakoid Membranes

The photo-stability of photosystem I (PSI) is of high importance for the photosynthetic processes. For this reason, we studied the protective action of two biogenic polyamines (PAs) spermine (Spm) and spermidine (Spd) on PSI activity in isolated thylakoid membranes subjected to photoinhibition. Our results show that pre-loading thylakoid membranes with Spm and Spd reduced considerably the inhibition of O₂ uptake rates, P700 photooxidation and the accumulation of superoxide anions (O₂ ⁻) induced by light stress. Spm seems to be more effective than Spd in preserving PSI photo-stability. The correlation of the extent of PSI protection, photosystem II (PSII) inhibition and O₂ ⁻ generation with increasing Spm doses revealed that PSI photo-protection is assumed by two mechanisms depending on the PAs concentration. Given their antioxidant character, PAs scavenge directly the O₂ ⁻ generated in thylakoid membranes at physiological concentration (1 mM). However, for non-physiological concentration, the ability of PAs to protect PSI is due to their inhibitory effect on PSII electron transfer.     

Amelioration of salt-induced oxidative stress in eggplant by application of 24-epibrassinolide

The effects of exogenous 24-epibrassinolide (EBR) on the growth, oxidative damage, antioxidant system and ion contents in eggplant (Solanum melongena L.) seedlings under salt stress were investigated. Eggplant seedlings were exposed to 90 mM NaCl with 0, 0.025, 0.05, 0.10 and 0.20 mg dm⁻³ EBR for 10 d. EBR, especially at concentration 0.05 mg dm⁻³, alleviated growth suppression caused by NaCl stress, decreased electrolyte leakage, superoxide production and content of malondialdehyde and H₂O₂ in NaCl-treated plants. EBR also increased activities of superoxide dismutase, guaiacol peroxidase, catalase and ascorbate peroxidase and the contents of ascorbic acid and reduced glutathione. Furthermore, we also found that Na⁺, Cl⁻ contents were decreased, K⁺, Ca²⁺ contents and K⁺/Na⁺, Ca²⁺/Na⁺ ratios were increased in the presence of EBR under salt stress.     

Response of Strawberry Plant cv. ‘Camarosa’ to Salicylic Acid and Methyl Jasmonate Application Under Salt Stress Condition

The possible role of salicylic acid (SA) and methyl jasmonate (MJ) treatments on the physiology responses and growth of strawberry (Fragaria?×?ananassa) cv. ‘Camarosa’ subjected to the different levels of salinity stress were investigated. Root and shoot growth as well as their Na⁺/K⁺ ratio, photosynthetic-related factors, and activity of some important antioxidant enzymes were determined in the salt-treated plants. Results indicated that salt stress reduced plant performance especially at higher concentrations. By increasing the levels of salinity stress, fresh and dry weight of shoot and roots, net photosynthetic rate (Pn), and stomatal conductance (Gs) significantly decreased, whereas intercellular CO₂ (Ci) increased. Application of exogenous SA and MJ significantly improved the plant physiological characters as well as fresh and dry weight of shoots and roots. Moreover, the ratio of Na⁺/K⁺ was elevated in the leaves and roots concomitantly with salinity levels, whereas SA and MJ treatments significantly reduced this ratio. Results of enzymatic assays showed that activity of ascorbate peroxidase, peroxidase, and superoxide dismutase enzymes increased in the salt-stressed plants. In addition, SA and MJ treatments reduced the destructive effects of salinity in strawberry plant. In general, among the tested concentrations, 0.5 mM SA and 0.25 mM MJ best increased the activity of antioxidant enzymes and hence alleviated the detrimental effects of salinity stress.     

Role of spermidine and spermine in alleviation of drought-induced oxidative stress and photosynthetic inhibition in Chinese dwarf cherry (Cerasus humilis) seedlings

To investigate the effect of exogenous Spermidine (Spd) and Spermine (Spm) on drought-induced damage to seedlings of Cerasus humili, relative water content (RWC), malondialdehyde content, relative electrolyte leakage, superoxide (O₂ ^?, SOD) generation rate, hydrogen peroxide (H₂O₂), endogenous polyamines (PAs), antioxidant enzymes [SOD and peroxidase (POD)] activities, PA-biosynthetic enzymes [arginine decarboxylase (ADC), ornithine decarboxylase (ODC), S-adenosylmethionine decarboxylase (SAMDC)] activities, as well as photosynthetic parameters, were measured in greenhouse cultured seedlings of C. humili. The results showed that either exogenous Spd or Spm (0.2 mM) significantly enhanced the level of RWC and prevented drought-induced lipid peroxidation. They also significantly enhanced photosynthetic capability and decreased O₂ ^? generation rate and H₂O₂ content. In addition, Spd and Spm helped to maintain SOD and POD activities in C. humili seedlings subjected to water stress, suggesting that they exerted a positive effect on antioxidant systems. The contents of endogenous free putrescine, Spd and Spm were increased to different extents in water-stressed C. humili seedlings. By the end of drought treatment (21 days) with exogenous Spd or Spm, the contents of free Spd increased by 30 and 38 %, respectively, and endogenous Spm increased by 41 and 26 %, respectively, compared with water-stressed plants. Furthermore, exogenous Spd or Spm enhanced the activities of ADC, ODC, and SAMDC. The pretreatment with Spd or Spm prevents oxidative damage induced by drought, and the protective effect of Spd was found to be greater than that of Spm.     

Effect of varying NaCl doses on flavonoid production in suspension cells of Ginkgo biloba: relationship to chlorophyll fluorescence,ion homeostasis,antioxidant system and ultrastructure

Ginkgo suspension cells were used to investigate the mechanism that governs the shift between primary and secondary metabolism under NaCl elicitation. The production of three flavonol glycosides, chlorophyll fluorescence, ion content, the antioxidant system, and the cellular ultrastructure in the presence of NaCl doses from 5 to 175 mM were examined. At low salt doses (5–50 mM), cell growth and flavonol glycosides accumulation were stimulated without damaging cell structure or inducing oxidative stress by maintaining high K⁺ and chlorophyll content. At moderate salt doses (75–125 mM), the cells could withstand the salt stress without an impact on survival by changing internal cellular structure, maintaining high levels of K⁺ and Ca²⁺ and increasing anti-oxidative enzyme activities rather than flavonol glycosides to counteract the inhibition of the photosystem II, the accumulation of Na⁺ and hydrogen peroxide (H₂O₂) in the cells. This allowed cells to divert their metabolism from growth to defense-related pathways and tolerate NaCl stress. At higher salinity (150–175 mM), the cellular structure was damaged, and the high Na⁺ and low K⁺ content led to osmotic stress, and therefore, the stimulation of peroxidase (POD) and catalase (CAT) was not enough to cope with high H₂O₂ accumulation. The high production of flavonol glycosides may be a response of elicitation stimulation to serious damage at 175 mM NaCl. In conclusion, the use of 175 mM NaCl may be desirable for the induction of flavonol glycoside production in Ginkgo suspension cells.