Long term exogenous putrescine application improves grain yield of a salt-sensitive rice cultivar exposed to NaCl

In order to determine the impact of polyamines on the yield-related parameters of rice (Oryza sativa L.) exposed to NaCl, the plants belonging to a salt-sensitive cultivar I Kong Pao were maintained from the young seedling stage until harvest on nutrient solutions containing 0 or 30 mM NaCl in the presence or absence of 10 μM putrescine (Put), 10 μM spermidine (Spd) or 10 μM spermine (Spm). Exogenous Put and to a lesser extent exogenous Spd improved growth and yield of salt-treated plants in relation to an increase in K⁺/Na⁺ ratio of shoots and roots as compared to plants exposed to NaCl in the absence of exogenous polyamines. Exogenous Put also improved the net CO₂ assimilation, at least partly as a consequence of an increase in the stomatal conductance. Yield increase of salt-treated plants exposed to Put was related to an improvement of floral morphogenesis leading to a higher number of fertile tillers per plant and a higher number of spikelets per panicle. Putrescine also improved the pollen viability in salt-treated plants, allowing a higher seed set and thus a higher grain yield per plant. Although polyamines accumulated in the shoots to some extent in response to exogenous application, neither Put nor Spd accumulated in the seeds. In contrast, Spm did not afford any protection of salt-treated plants but was translocated to the seeds during maturation. Seeds with a high internal Spm concentration exhibited delayed germination in the presence of NaCl. These data are discussed in relation to the implication of polyamine in the metabolism and physiology of salt-treated plants.     

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.     

Exogenous application of free polyamines enhance salt tolerance of pistachio (Pistacia vera L.) seedlings

The protective effects of free polyamines (PAs) against salinity stress were investigated for pistachio seedlings (Pistacia vera cv. Badami-Zarand) in a controlled greenhouse. Seedlings were treated with 25, 50, 100 and 150 mM of salts including NaCl, CaCl₂ and MgCl₂. Foliar treatments of putrescine, spermidine (Spd) and spermine (Spm) (0.1 and 1 mM) were applied during the salinity period. Results showed that growth characteristics of pistachio seedlings decreased under salinity stress and the application of PAs efficiently reduced the adverse effects of salt stress. PAs reduced the severe effects of salt stress in pistachio seedlings neither by increasing the activities of peroxidase and ascorbate peroxidase nor by increasing the proline content but by increasing the activities of superoxide dismutase and catalase and decreasing the hydrogen peroxide (H₂O₂) activity. PAs treated seedlings showed a lower Na⁺:K⁺ ratio and Cl^? in leaves suggesting the role of PAs in balancing the ion exchange and better Na⁺:K⁺ discrimination under salt stress condition. These results showed the promising potential use of PAs especially Spm and Spd for reducing the negative effects of salinity stress and improving the growth of pistachio seedlings.     

Effect of NaCl and Polyamines on Plasma Membrane Lipids of Wheat Roots

Caryopses of a salt sensitive wheat cultivar (Triticum aestivum L. cv. Giza 163) were presoaked in 2.5 mM putrescine (Put), 5 mM spermidine (Spd) or 2.5 mM spermine (Spm) for 24 h and then subjected to 150 mM NaCl added to the growth medium for 15 d. Effects of NaCl and polyamines (PAs) on plasma membrane (PM) lipids, phospholipids, fatty acids, and free sterols were determined. NaCl treatment caused a decrease in total phospholipids, increase in saturated fatty acids and altered distribution of sterols and phospholipids. NaCl also induced increase in sterol/phospholipid ratio. PAs treatments (particularly Put and Spd) counterbalanced the NaCl deleterious effects on PM lipids.     

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).     

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.     

Do exogenous polyamines have an impact on the response of a salt-sensitive rice cultivar to NaCl?

In order to analyze the putative impact of polyamines (PAs) on the plant response to salt, seedlings from the salt-sensitive rice cultivar I Kong Pao (IKP) were exposed for 5, 12 and 19 days to 0, 50 or 100 mM NaCl in the absence, or in the presence of exogenous PAs (putrescine (Put), spermidine (Spd) or spermine (Spm) 1mM) or inhibitors of PA synthesis (methylglyoxalbis-guanyl hydrazone (MGBG) 1mM, cyclohexylammonium (CHA) 5mM and D-arginine (D-Arg) 5mM). The addition of PAs in nutritive solution reduced plant growth in the absence of NaCl and did not afford protection in the presence of salt. PA-treated plants exhibited a higher K+/Na+ ratio in the shoots, suggesting an improved discrimination among monovalent cations at the root level, especially at the sites of xylem loading. The diamine Put induced a decrease in the shoot water content in the presence of NaCl, while Spd and Spm had no effects on the plant water status. In contrast to Spd, Spm was efficiently translocated to the shoots. Both PAs (Spd and Spm) induced a decrease in cell membrane stability as suggested by a strong increase in malondialdehyde content of PA-treated plants exposed to NaCl. These results are discussed in relation to the putative functions of PAs in stressed plant metabolism.     

Effect of exogenous spermidine on polyamine metabolism in water hyacinth leaves under mercury stress

The influence of exogenous spermidine (Spd) on arginine decarboxylase (ADC), ornithine decarboxylase (ODC), polyamine oxidase (PAO) activities and polyamines (PAs), proline contents in water hyacinth leaves under Mercury (Hg) stress was investigated after 6 days treatment. The results showed that free putrescine (Put) content increased, the contents of free spermidine (Spd) and spermine (Spm) and the (Spd + Spm)/Put ratio in water hyacinth leaves decreased significantly with the increase of the Hg concentrations. Hg stress also disturbed the activities of ADC, ODC and PAO and caused changes on proline content. Compared to the Hg-treatment only, exogenous Spd (0.1 mM) significantly reduced the accumulation of free Put, increased the contents of free Spd and Spm and the ratio of (Spd + Spm)/Put in water hyacinth leaves. Furthermore, exogenous Spd enhanced the activities of ADC, ODC and PAO and significantly increased proline content. The PS-conjugated PAs and PIS-bound PAs changed in the same trend as free PAs. These results suggest that exogenous Spd can alleviate the metabolic disturbance of polyamines caused by Hg in water hyacinth leaves.     

氯化钠胁迫对南瓜根系游离态多胺含量和活性氧水平的影响

以中国南瓜杂交种‘360.3×112.2’和黑籽南瓜为试验材料,在营养液栽培条件下研究了NaCl胁迫对两种南瓜植株生长、根系活性氧水平和游离态多胺含量的影响.结果表明,NaCl胁迫10 d后,与对照相比,两种南瓜植株生长都受到明显抑制,但中国南瓜杂交种比黑籽南瓜植株的耐盐性强.NaCl胁迫使南瓜根系O₂^-·产生速率和H₂O₂含量提高,且黑籽南瓜的O₂^-·产生速率和H₂O₂含量高于中国南瓜杂交种.两种南瓜根系中腐胺(Put)、亚精胺(Spd)、精胺(Spm)和多胺(PAs)含量及Put/PAs高于对照,并呈现先升后降的趋势;根系中（Spd+Spm）/Put低于对照,呈现先降后升的趋势.中国南瓜杂交种根系中Put含量和Put/PAs低于黑籽南瓜,而Spd、Spm含量和（Spd+Spm）/Put高于黑籽南瓜.表明两种南瓜根系中多胺含量的升高对减少或清除组织中的活性氧有积极作用,Put向Spd、Spm的转化有利于增强植株的耐盐性;中国南瓜杂交种‘360.3×112.2’的耐盐性高于黑籽南瓜与其根系中Put/PAs较低、（Spd+Spm）/Put和PAs含量较高,使其清除活性氧能力较强有关.     

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.     

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.     

Selection of callus cultures of sugarcane (Saccharum sp.) tolerant to NaCl and their response to salt stress

Stable callus cultures tolerant to NaCl (68 mM) were developed from salt-sensitive sugarcane cultivar CP65-357 by in vitro selection process. The accumulation of both inorganic (Na⁺, Cl⁻ and K⁺) and organic (proline and soluble sugars) solutes was determined in selected and non-selected calli after a NaCl shock in order to evaluate their implication in in vitro salt tolerance of the selected lines. Both salt-tolerant and non-selected calli showed similar relative fresh weight growth in the absence of NaCl. No growth reduction was observed in salt-tolerant calli while a significant reduction about 32% was observed in nonselected ones when both were cultivated on 68 mM NaCl. Accumulation of Na⁺ was similar in both salt-tolerant and non-selected calli in the presence of NaCl. Accumulation of Cl⁻ was lower in NaCl-tolerant than in non-selected calli while proline and soluble sugars were more accumulated in salt-tolerant than in non-selected calli when both were exposed to salt. K⁺ level decreased more severely in non-selected calli than in NaCl-tolerant ones after NaCl shock. The results indicated that K⁺ and Cl⁻ may play a key role in in vitro salt-tolerance in sugarcance cell lines obtained by in vitro selection and that organic solutes could contribute mainly to counteract the negative water potential of the outside medium.     

多胺对宁夏枸杞愈伤组织器官发生和体细胞胚发生的影响

利用已建立的宁夏枸杞（Lycium barbarumL.)愈伤组织器官发生和体细胞胚发生体系,对多胺在其离体形态发生中的作用进行研究。通过检测内源多胺含量发现,在所研究的三种多胺中,Put是器官发生途径的主要多胺,而在体细胞胚发生途径Spd含量占优势。Put含量变化在两条途径中相似：在愈伤组织分化的早期迅速积累不仅又下降,随着芽原基和球形胚的形成含量又进一步上升。器官发生中Spd最高含量仅在培养的第一天后,Spd含量才开始上升,到第十天时达到最高值。三种外源多胺的添加均有有效地促进两种离体分化途径的形态建成：Spd(100 μmol/L）能显著增加不定芽数,而体细胞胚发生中Spd(100μmol/L）而Put（100μmol/L）的单独处理能最好地促进体细胞胚形成和进一步发育成苗;尽管Spm在离体形态发生中含量较低,但添加外源Spm也促进了不定芽形成和体细胞胚形成然后成苗。多胺生物合成抑制剂CHA处理阻碍不定芽形成和和体细胞胚的进一步发育;但是MGBG对器官发生途径中的形态建成没有影响,即降低体细胞胚的发生频率及再生苗数。添加Spd(50μmol/L)能部分逆转CHA、MGBG的抑制效应。以上结果表明,多胺对宁夏枸杞器官发生和体细胞胚发生途径的离体形态建成有一定影响。     

NaCl胁迫对菜用大豆种子多胺代谢的影响

采用蛭石栽培,在100 mmol·L-1NaCl胁迫下,对耐盐性不同的两个品种菜用大豆种子的丙二醛(MDA)含量和多胺(PAs)代谢进行了研究.结果表明:NaCl胁迫显著增加了菜用大豆种子的MDA含量,但耐盐品种‘绿领特早’(LL)的增幅低于盐敏感品种‘理想高产95-1’(LX).与LX相比,LL种子在整个NaCl胁迫期间均维持了相对较高的游离态精胺(Spm)、结合态Spm、结合态亚精胺(Spd)、束缚态Spd和束缚态腐胺(Put)含量,较高的(Spd +Spm )/Put 和(cPAs+bPAs)/fPAs值及较低的Put/PAs值,在胁迫中、后期(9～15 d)维持了相对较高的游离态Spd含量;胁迫期间,LL的精胺酸脱羧酶(ADC)长时期(6～15 d)保持相对较高的活性,而多胺氧化酶(PAO)则长时期(6～15 d)维持相对较低的活性.综上,LL具有较强的多胺合成能力及较强的Put向Spd和Spm以及游离态多胺向结合态和束缚态多胺转化的能力,进而有效抑制了细胞的膜脂过氧化,这可能是其耐盐性较强的重要原因之一.     

Exogenous polyamines enhance copper tolerance of Nymphoides peltatum

The protective effects of polyamines (PAs) against copper (Cu) toxicity were investigated in the leaves of Nymphoides peltatum. Cu treatment increased the putrescine (Put) level and lowered spermidine (Spd) and spermine (Spm) levels, thereby reducing the (Spd+Spm)/Put ratio in leaves. Exogenous application of Spd or Spm markedly reversed these Cu-induced effects for all three PAs and partially restored the (Spd+Spm)/Put ratio in leaves. It also significantly enhanced the level of proline, retarded the loss of soluble protein, decreased the rate of O2*- generation and H2O2 content, and prevented Cu-induced lipid peroxidation. Furthermore, exogenous Spd and Spm reduced the accumulation of Cu and effectively maintained the balance of nutrient elements in plant leaves under Cu stress. These results suggest that exogenous application of Spd or Spm can enhance the tolerance of N. peltatum to Cu by increasing the levels of endogenous Spd and Spm as well as the (Spd+Spm)/Put ratio.     

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.     

Stress-dependent accumulation of spermidine and spermine in the halophyte Mesembryanthemum crystallinum under salinity conditions

We studied the effects of chloride salinity (300 and 500 mM NaCl) on the content of free polyamines (PAs) from putrescine (Put) family in Mesembryanthemum crystallinum L. leaves and roots. The contents of Put and spermidine (Spd) in leaves increased temporarily, achieving the highest values on the third day of salinity treatment; thereafter (by days 7–14), they dropped sharply. The content of spermine (Spm) increased gradually, and its high level was maintained until the end of experiment. The dynamics of Spm accumulation in leaves under salinity conditions resembled that of phosphoenolpyruvate carboxylase (PEPC), a key enzyme of the water-saving CAM pathway of photosynthesis. This indicates indirectly the involvement of Spm in the common ice plant adaptation to salinity. A decrease in the molar ratios of Spd to Spm in the leaves under salinity conditions could point to the acceleration of Spm biosynthesis (accumulation) during plant adaptation, whereas the levels of Spm precursors, Put and Spd, did not increase. This phenomenon could be explained by an accelerated conversion of Spd into Spm, an active liberation of free Spm from its conjugates, or changes in the rates of studied PA biosynthesis and degradation under salinity. At the same time, the intracellular concentration of ethylene rose under these conditions. It was supposed and then demonstrated, that the pathway of ethylene biosynthesis and that of the synthesis of Put family PAs compete under severe salinity conditions. This competition might be based on the disturbances in sulfur metabolism and a decrease in the methionine content, an immediate precursor of S-adenosyl-L-methionine.     

Polyamines content and physiological and biochemical responses to ladder concentration of nickel stress in Hydrocharis dubia (Bl.) Backer leaves

Influence of ladder concentration of nickel (Ni) on the leaves of Hydrocharis dubia were studied after 3 days treatment. The accumulation of Ni, the content of polyamines, proline, malondialdehyde (MDA) and soluble protein, as well as the activities of superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD) in the leaves were investigated. The result indicated that the toxicity of Ni manifested in respective aspect of physiological and biochemical characters. Significant increase of Ni concentration in the leaf tissue was observed, which was concentration dependent. Visible symptoms of Ni toxicity: chlorosis and necrosis occurred following the 3rd day. Meantime, treatment with Ni resulted in the increase in the generation rate of O₂^•− in the leaves. SOD and CAT activities decreased significantly in response to Ni treatment, it was possibly the reason of accumulation of O₂^•−. However, a several-fold decrease in POD activities was found. Our results indicated that because of prolonged increases in O₂^•− level, oxidative damage, measured as the level of lipid peroxidation, occured in the leaves of Ni treated fronds. The changes of the content of polyamines (PAs) were also investigated in the leaves of Hydrocharis dubia. Ni treatment significantly increased the putrescine (Put) level and lowered spermidine (Spd) and spermine (Spm) levels, thereby significantly reducing the ratio of free (Spd + Spm)/Put in leaves, which has been considered as the signal under stress. Although the trend that PS-conjugated PAs and PIS-bound PAs changed the same as free PAs, they changed in more less extent.     

Effect of polyamines on in vitro somatic embryogenesis in <Emphasis Type="Italic">Momordica</Emphasis><Emphasis Type="Italic">charantia</Emphasis> L.

The effects of exogenous polyamines (PAs) on enhancement of somatic embryogenic calli was investigated in Momordica charantia L. in vitro. Induction of somatic embryogenesis (SE) in leaf explants of M. charantia after 21 days of culture in Murashige and Skoog (MS) medium was determined using scanning electron microscopy. During induction of SE there were high titers of Putrescine (Put) as compared to Spermidine (Spd) and Spermine (Spm), a prerequisite for cell division. Addition of PAs to the embryogenic media resulted in an increase in fresh weights and number of somatic embryos of 21-day old embryogenic calli. Put at a concentration of 1 mM showed maximum increase in fresh weights of embryogenic calli (5 fold) and number of somatic embryos produced per 0.2 g of callus (2.5 fold). Moreover addition of PAs to the embryogenic media resulted in lowering of endogenous free PA level of 21-day old embryogenic calli. Thus, when the media was supplemented with exogenous PAs a positive correlation was found to exist between Somatic Embryogenesis enhancement and decrease in endogenous free PA levels.     

Polyamine and nitric oxide levels relate with morphogenetic evolution in somatic embryogenesis of <Emphasis Type="Italic">Ocotea catharinensis</Emphasis>

In this study we examined the effect of polyamines (PAs) putrescine (Put), spermidine (Spd) and spermine (Spm) on growth, morphology evolution, endogenous PAs levels and nitric oxide (NO) release in Ocotea catharinensis somatic embryo cultures. We observed that Spd and Spm reduced culture growth, permitted embryo morphogenetic evolution from the earliest to last embryo development stages, increased endogenous PAs levels, and induced NO release in O. catharinensis somatic embryos. On the other hand, Put had little effect on these parameters. Spd and Spm could successfully be used to promote somatic embryo maturation in O. catharinensis. The results suggest that Spd and Spm have an important role during the growth, development and morphogenetic evolution of somatic embryos, through alterations in the endogenous nitric oxide and PAs metabolism in this species.