Plasticity of polyamine metabolism associated with high osmotic stress in rape leaf discs and with ethylene treatment

In rape leaf discs the response to osmotic stress has been found to be associated with increases in putrescine and 1,3-diaminopropane (an oxidation product of spermidine and/or spermine) and decreases in spermidine titers. In contrast, agmatine and spermine titers showed small changes while cadaverine accumulated massively. Similar results were observed in whole rape seedlings subjected to drought conditions. -DL-difluoromethylarginine (DFMA), a specific irreversible inhibitor of arginine decarboxylase, strongly inhibited polyamine accumulation in unstressed rape leaf discs, which suggested that the arginine decarboxylase pathway is constitutively involved in putrescine biosynthesis. In leaf discs treated under high osmotic stress conditions, both DFMA and DFMO (-DL-difluoromethylornithine, a specific and irreversible inhibitor of ornithine decarboxylase) inhibited the accumulation of polyamines. Although the stressed discs treated with DFMA had a lower concentration of putrescine than those treated with DFMO, we propose that under osmotic stress the synthesis of putrescine might involve both enzymes. DFMA, but not DFMO, was also found to inhibit cadaverine formation strongly in stressed explants. The effects on polyamine biosynthesis and catabolism of cyclohexylamine, the spermidine synthase inhibitor, aminoguanidine, the diamine-oxidase inhibitor and -aminobutyric acid, a product of putrescine oxidation via diamine oxidase or spermidine oxidation via polyamine oxidase were found to depend on environmental osmotic challenges. Thus, it appears that high osmotic stress did not block spermidine biosynthesis, but induced a stimulation of spermidine oxidation. We have also demonstrated that in stressed leaf discs, exogenous ethylene, applied in the form of (2-chloroethyl) phosphonic acid or ethephon, behaves as an inhibitor of polyamine synthesis with the exception of agmatine and diaminopropane. In addition, in stressed tissues, when ethylene synthesis was inhibited by aminooxyacetic acid or aminoethoxyvinylglycine, S-adenosylmethionine utilization in polyamine synthesis was not promoted. The relationships between polyamine and ethylene biosynthesis in unstressed and stressed tissues are discussed.     

Polyamines and related enzymes in rice seeds differing in germination potential

In ungerminated rice seeds, (Japonica rice variety, CV Tapei 309), the content of free amines (putrescine, spermidine, spermine, tyramine) was higher in seed lots having a low germination frequency compared to those with high germination potential. Conversely, amine conjugates (di-feruloylputrescine, di-feruloylspermidine, diferuloyldiaminopropane and feruloyltyramine) decreased with loss of viability. Thus, these compounds appeared to constitute biochemical markers of seed viability. In seeds with high germination potential, conjugates decreased drastically during germination, with an early and rapid increase in free amines (putrescine, spermidine, tyramine). Arginine decarboxylase (ADC) activity was highest during the germination of high germination potential seeds, its activity gradually declining with loss of viability and being closely correlated with agmatine content. The polyamine biosynthetic inhibitors (-DL-difluoromethylarginine, DFMA, a specific and irreversible inhibitor of ADC; -DL-difluoromethylornithine, DFMO, a specific irreversible inhibitor of ornithine decarboxylase (ODC); cyclohexylammonium sulfate, CHA, inhibitor of spermidine synthase) neither depleted putrescine and spermidine levels nor inhibited germination in high germination potential seeds. In low germination potential seeds, the germination process was inhibited by DFMA or CHA. Application of agmatine resulted in a reversal of inhibition. DFMA inhibited ADC activity in both categories of seeds. In low germination potential seeds treated with CHA no ADC activity was found. These results suggest that amines are involved in the germination process of rice seeds. It appears that amine conjugates may serve as a storage form of amines which, upon enzymatic hydrolysis, could supply the cell with an additional amine reserve and influence cell division and/or cell elongation.Abbreviations ADC arginine decarboxylase - ODC ornithine decarboxylase - DFMA -DL-difluoromethylarginine - DFMO -DL-difluoromethylornithine - CHA cyclohexylammonium sulfate     

Effect of salt and osmotic stress on changes in polyamine content and arginine decarboxylase activity in Lupinus luteus seedlings

The effects of NaCl (260 mM) and sorbitol (360 mM) isoosmotic stresses on polyamine titers in lupin (Lupinus luteus L. var. Ventus) in relation to organ-specific responses were investigated. Analysis showed that during the first few hours (4 h) of salt and osmotic stress higher amounts of putrescine (Put) and spermidine (Spd) were accumulated in the roots and leaves of lupin seedlings. After exposing the plants to a longer duration (24 h) of exposure to NaCl, the level of free Put decreased in roots and cotyledons by about 48% and 54%, respectively, and increased in hypocotyls and leaves by about 27% and 73%, respectively. The Level of free Spd also decreased in roots by about 50%, in contrast to the increase of Spd observed in hypocotyls and leaves by about 50% and 70%, respectively. The effect of non-ionic stress on the level of Put and Spd in studied organs of lupin was similar to that of NaCl. Free spermine was at an undetectable level in examined organs. However, in the roots of lupin growing for 24 h in the presence of NaCl and/or sorbitol, the activity of arginine decarboxylase (ADC) (EC 4.1.1.19) increased by about 66% and 80%, respectively. ADC activity in leaves was similar to that observed in the control. Additionally, in the roots and leaves of lupin growing under the stress condition (NaCl or sorbitol), a higher level of polyamines (PAs) bound to microsomal membranes was observed. It is probable that PAs bound to microsomal membranes prevent stress-induced damage. We conclude that both stresses induce biosynthesis of Put and other PAs in the roots, as well as Put accumulation in the leaves, and this may indicate translocation of Put from the roots to the shoot. The possible role of PAs in adaptive mechanisms to stress is discussed.     

Exogenously applied polyamines increase drought tolerance of rice by improving leaf water status,photosynthesis and membrane properties

Drought stress hampers rice performance principally by disrupting the plant–water relations and structure of biological membranes. This study appraised the role of polyamines (PAs) in improving drought tolerance in fine grain aromatic rice (Oryza sativa L.). Three PAs [putrescine (Put), spermidine (Spd) and spermine (Spm)] were used each at 10 μM as seed priming (by soaking seeds in solution) and foliar spray. Primed and non-primed seeds were sown in plastic pots with normal irrigation in a phytotron. At four-leaf stage, plants were subjected to drought stress by bringing the soil moisture down to 50% of field capacity by halting water supply. For foliar application, 10 μM solutions each of Put, Spd and Spm were sprayed at five-leaf stage. Results revealed that drought stress severely reduced the rice fresh and dry weights, while PAs application improved net photosynthesis, water use efficiency, leaf water status, production of free proline, anthocyanins and soluble phenolics and improved membrane properties. PAs improved drought tolerance in terms of dry matter yield and net photosynthesis was associated with the maintenance of leaf water status and improved water use efficiency. Among the antioxidants, catalase activity was negatively related to H₂O₂ and membrane permeability, which indicated alleviation of oxidative damage on cellular membranes by PAs application. Foliar application was more effective than the seed priming, and among the PAs, Spm was the most effective in improving drought tolerance.     

Polyamines in grapevine microcuttings cultivated in vitro. Effects of amines and inhibitors of polyamine biosynthesis on polyamine levels and microcutting growth and development

The main free amines identified during growth and development of grapevine microcuttings of rootstock 41 B, (Vitis vinifera cv. Chasselas × Vitis berlandieri) cultivated in vitro were agmatine, putrescine, spermidine, spermine, diaminopropane and tyramine (an aromatic amine). Amine composition differed according to tissue, with diaminopropane the major polyamine in the apical parts, internodes and leaves. Putrescine predominated in the roots. There was also a decreasing general polyamine and specific tyramine gradient along the stem from the top to the bottom. Conjugated amines were only found in roots. The application of exogenous amines (agmatine, putrescine, spermidine, tyramine) stimulated development and growth of microcuttings, suggesting that the endogenous concentrations of these amines can be growth limiting. Diaminopropane (the product of oxidation of spermidine or spermine by polyamine oxydases) strongly inhibited microcutting growth and development. -DL-difluoromethylarginine (DFMA), a specific and irreversible inhibitor of the putrescine-synthesizing enzyme, arginine decarboxylase (ADC), led to inhibition of microcutting development. Application of agmatine or putrescine to the inhibited system resulted in a reversal of inhibition indicating that polyamines are involved in regulating the growth and development of grapevine microcuttings. -DL-difluoromethylornithine (DFMO), a specific and irreversible inhibitor of putrescine biosynthesis from ornithine decarboxylase (ODC), had no effect on microcutting development and growth. We propose that ADC regulates putrescine biosynthesis during microcutting development.     

Polyamine Metabolism and Osmotic Stress : II. Improvement of Oat Protoplasts by an Inhibitor of Arginine Decarboxylase

We have attempted to improve the viability of cereal mesophyll protoplasts by pretreatment of leaves with dl-α-difluoromethylarginine (DFMA), a specific `suicide' inhibitor of the enzyme (arginine decarboxylase) responsible for their osmotically induced putrescine accumulation. Leaf pretreatment with DFMA before a 6 hour osmotic shock caused a 45% decrease of putrescine and a 2-fold increase of spermine titer. After 136 hours of osmotic stress, putrescine titer in DFMA-pretreated leaves increased by only 50%, but spermidine and spermine titers increased dramatically by 3.2- and 6-fold, respectively. These increases in higher polyamines could account for the reduced chlorophyll loss and enhanced ability of pretreated leaves to incorporate tritiated thymidine, uridine, and leucine into macromolecules. Pretreatment with DFMA significantly improved the overall viability of the protoplasts isolated from these leaves. The results support the view that the osmotically induced rise in putrescine and blockage of its conversion to higher polyamines may contribute to the lack of sustained cell division in cereal mesophyll protoplasts, although other undefined factors must also play a major role.     

Senescence of Rice Leaves XXX Levels of Endogenous Polyamines and Dark-Induced Senescence of Rice Leaves

The role of endogenous polyamines in the control of dark-inducedsenescence of detached rice leaves was investigated by quantitatinglevels of various polyamines by HPLC. Putrescine, spermidineand spermine were all present throughout senescence. Neithercadaverine nor 1,3-diaminopropane was detected. During dark-inducedsenescence, there was a marked decrease in levels of putrescineand an increase in those of spermidine and spermine. The rateof production of ethylene increased markedly upon excision ofleaves. -Difluoromethylarginine (DFMA) and -difluoromethylornithine(DFMO) caused a reduction in levels of putrescine, yet had noeffect on levels of spermidine and spermine. Neither DFMA norDFMO had any effect on senescence or on the production of ethylene.Treatment with dicyclohexylamine (DCH) and methylglyoxal bis-(guanylhydrazone)(MGBG) reduced levels of spermine and increased those of putrescinein detached leaves. After treatment with DCH or MGBG, both senescenceand the production of ethylene were significantly promoted.The current results suggest that endogenous polyamines may notplay a significant role in the control of dark-induced senescenceof rice leaves. This conclusion is supported by the furtherobservations that (a) benzyladenine, which is known to retardsenescence, decreased levels of putrescine but had no effecton those of spermidine and spermine; and (b) ABA, which promotedsenescence, increased levels of putrescine and had no effecton those of spermidine and spermine. (Received March 30, 1991; Accepted June 27, 1991)     

Changes in polyamine concentration during seed germination

Phaseolus mungo seeds 0 to 10 days after germination contained putrescine, spermidine, spermine, cadaverine, agmatine and tyramine. The rate of biosynthesis of total polyamines, proteins and RNA in the developing seeds follows similar profiles, reaching maxima 3 hr from germination. Putrescine, cadaverine, spermidine, spermine and agmatine were the major amines found in Pisum sativum 0–7 days after germination. RNA and proteins seem to follow the same pattern as polyamines during the first 12 hr in the developing pea seeds. RNA reaches a peak at 15 hr and polyamines and proteins peak 24 hr after germination. A rise to total polyamine concentration was also observed in seeds of Tragopogon porrifolius, Zea mays and Triticum aestivum 2–12 hr after germination.     

Changes in chlorophyll,polyamines and chloroplast ultrastructure of Puccinia striiformis induced ‘green islands’ on detached leaves of Triticum aestivum

Biochemical and ultrastructure features of ‘green islands’ were investigated using detached wheat leaves infected with the yellow rust Puccinia striiformis. Chlorophylls appear to culminate 10 d after inoculation at which point ‘green islands’ were fully developed. These changes were paralleled by an increase in spermidine and spermine content which play an important role in formation of ‘green islands’. Retention of chlorophyll was demonstrated in leaf tissues of wheat plants supplied with exogenous putrescine, spermidine and spermine. Putrescine was least and spermidine and spermine most effective in retarding loss of chlorophylls. Ultrastructural observation revealed that chloroplasts were regenerated in ‘green islands’ where many proplastids were detected. The regeneration of chloroplasts coincided with the high concentration of chlorophylls and polyamines particularly spermidine and spermine. The ultrastructural changes of chloroplasts in leaf cell containing infection structures were parallel to physiological changes.     

Relationships between polyamines, ethylene, osmoprotectants and antioxidant enzymes activities in wheat seedlings after short-term PEG- and NaCl-induced stresses

Contents of ethylene, osmoprotectants, levels and forms of polyamines (PAs) and activities of antioxidant enzymes in the leaves and roots were investigated for five wheat cultivar seedlings (differing in drought tolerance) exposed to osmotic stress (?1.5 MPa). Stress was induced by 2-day-long treatment of plants with polyethylene glycol 6000 (PEG) or NaCl added to hydroponic cultures. Nawra, Parabola and Manu cv. (drought tolerant) showed a marked increase in osmoprotectors (proline and soluble carbohydrates, mainly glucose, saccharose and maltose), free PAs (putrescine Put, spermidine Spd and spermine Spm) and Spd-conjugated levels, in both leaves and roots, after PEG-treatments. Radunia and Raweta (drought sensitive) exhibited smaller changes in the content of these substances. The analysis of enzymes involved in proline metabolism revealed the glutamate as a precursor of proline synthesis in PEG-induced stress conditions. The increase in the activity of antioxidative enzymes, especially catalase and peroxidases, was characteristic for tolerant wheat plants, but for sensitive ones, a decrease in superoxide dismutase and an increase in mainly glutathione reductase activities were observed. After NaCl-treatment smaller changes of all biochemical parameters were registered in comparison with PEG-induced stress. Exceptions were the higher values of ethylene content and a significant increase in saccharose, raffinose and maltose levels (only in stress sensitive plants). The proline synthesis pathway was stimulated from both glutamate and ornithine precursors. These results suggest that the accumulation of inorganic ions in NaCl-stressed plants may be involved in protective mechanisms as an additional osmoregultor. Thus, a weaker stressogenic effect as determined as water deficit by leaf relative water content and relative dry weight increase rate and differences in metabolite synthesis in comparison with PEG stress was observed. Proline seems to be the most important osmo-protector in osmotic stress initiated by both PEG and NaCl. The synthesis of sugars and PAs may be stimulated in a stronger stress conditions (PEG).     

Effect of mineral deficiency on amine formation in higher plants

Potassium deficiency caused putrescine accumulation in the leaves of barley, radish, pea, bean and spinach plants. Magnesium deficiency caused putrescine accumulation in barley, pea and bean leaves, and also in the leaves of older radish plants. In young radish plants less putrescine was found in magnesium deficiency, and in spinach magnesium deficiency was without effect on putrescine levels. Putrescine content may be a useful guide to the mineral status of legumes, since accumulation of this amine may be detected before deficiency symptoms appear. Radioactivity from l-arginine-[U-¹⁴C] fed to barley seedlings was detected in agmatine within 2 hr, and probably also in the hordatines after 24 hr, feeding. After 2 hr the label in the agmatine was greatest in the potassium-deficient plants, but after 24 hr the level declined to that found in the agmatine of the leaves of the magnesium-deficient and control seedlings. The rate of putrescine formation was high in both potassium and magnesium deficiency. Incorporation of radioactivity in spermidine and spermine on feeding putrescine-[1,4-¹⁴C] to barley seedlings was estimated in the dansylated amines after separation by TLC. Activity was higher in spermidine and lower in spermine in the potassium-deficient plants than in the controls. The spermidine/spermine ratio declined on excision of barley leaves.     

Interactive effect of cytokinin and potassium on sink-source relationships in Lupinus angustifolius

This study examined whether increased K supply in conjunction with BAPcould increase lupin seed yield and harvest index by enlarging sink volume (podnumber), increasing assimilate and improving assimilate partitioning to filltheadditional pods induced by BAP treatment. Narrow-leafed lupin(Lupinusangustifolius, cv. Danja abs^– mutant) was grown inaglasshouse, in pots containing sandy soil with four K treatments (0, 15, 60 and120 mg K/kg soil). BAP (2 mM) was applied daily toallmain stem flowers throughout the life of each flower from opening to senesced.BAP application did not affect assimilate production (as measured by totalabove-ground biomass), but changed assimilate partitioning. On BAP-treatedplants, there were greater proportions of seed to pod wall dry weight on themain stem but smaller proportions on the branches, and an increased weightratioof seed to pod wall overall which meant more assimilate was used for seedgrowthrather than pod wall growth. BAP increased the number of pods per plant by35% and this more than compensated for the decreases in seeds per podandseed weight. Therefore, there was an increased harvest index (+11%)and seed yield per plant (+13%) in BAP-treated plants. BAP alsoincreased the number of pods with filled seeds (146%) on the main stemand main stem seed K⁺ concentration (from 0.81% to0.87%). Added K increased biomass but only slightly affected assimilatepartitioning. As applied K increased, relatively more assimilate was used forpod wall growth rather than seed growth. Added K increased seed yield per plantby about 14% due to increases in seed weight and the number of pods onthe main stem. Moreover, K⁺ concentration in seeds and shootsincreased with increasing level of applied K. Seed yield was enhanced more byBAP when K was supplied at high levels. Increasing K supply interactedpositively with added BAP by increasing narrow-leaf lupin seed yield andharvestindex through increases in assimilate supply and its partitioning into seeds.     

Abscisic acid promotes accumulation of toxin ODAP in relation to free spermine level in grass pea seedlings (Lathyrus sativus L.).

Interrelationship among abscisic acid (ABA) content, accumulation of free polyamines and biosynthesis of beta-N-oxalyl-l-alpha,beta-diaminopropionic acid (ODAP) was studied in grass pea (Lathyrus sativus L.) seedlings under drought stress induced by 10% polyethylene glycol (PEG6000). Increase of ABA content occurred prior to that of ODAP and polyamine contents, and was found significantly positive correlation between ABA content and ODAP content. Addition of exogenous ABA increased ODAP content in leaves. On the other hand, pretreatment with alpha-difluoromethylarginine (DFMA), a polyamine biosynthesis inhibitor, significantly suppressed the accumulation of free putrescine (Put), free spermidine (Spd) and free spermine (Spm), which in turn inhibited biosynthesis of ODAP in well-watered leaves. Meanwhile, addition of exogenous Put alleviated DFMA-induced inhibition on the biosynthesis of Put and Spd, but did not affect the biosynthesis of Spm and ODAP in well-watered leaves. Same result was also achieved in drought-stressed leaves. Increasing accumulation of ODAP was significantly correlated with increasing Spm content (R=0.7957**) but not with that of Spd and Put. Therefore, it can be argued that ABA stimulated the biosynthesis of ODAP simultaneously with increasing the level of free Spm under drought stress condition.     

Changes in growth,photosynthetic capacity and ionic relations in spring wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.) due to pre-sowing seed treatment with polyamines

The influence of pre-sowing seed treatment with polyamines (2.5 mM putrescine, 5.0 mM spermidine and 2.5 mM spermine) on growth, photosynthetic capacity, and ion accumulation in two spring wheat (Triticum aestivum L.) cultivars MH-97 (intolerant) and Inqlab-91 (tolerant) was examined. The primed seeds of each treatment and non-primed seeds were sown in a field containing 15 dS m⁻¹ NaCl. Although all three polyamines were effective in improving shoot growth and grain yield in both cultivars under saline conditions, the effect of spermine was very pronounced particularly in improving grain yield. Different priming agents did not affect the net CO₂ assimilation rate and transpiration rate of either cultivar. However, pre-treatment with spermidine increased stomatal conductance (g_s) in the tolerant cultivar, whereas with spermine stomatal conductance decreased in the intolerant cultivar under salt stress. Priming agents had different effects on the accumulation of different ions in wheat plant tissues. When spermidine and distilled water were used as priming agents, they were effective in reducing shoot [Na⁺] in the tolerant and intolerant cultivars, respectively under saline conditions. Although all priming agents caused an increase in shoot [K⁺], distilled water was more effective in improving shoot [K⁺] in both cultivars under salt stress. Pre-treatment with spermidine was very effective in reducing shoot [Cl⁻] under saline conditions particularly in the tolerant cultivar. However, the pattern of accumulation of different ions in roots due to different seed priming treatments was not consistent in either cultivar except that root Na⁺ decreased due to priming with spermine and spermidine in the intolerant and tolerant cultivars under saline conditions. In conclusion, although all three priming agents, spermine, spermidine and putrescine, were effective in alleviating the adverse effect of salt stress on wheat plants, their effects on altering the concentration of different ions and growth were different in the two cultivars differing in salt tolerance.     

Effects of inhibitors of polyamine biosynthesis and of polyamines on strawberry microcutting growth and development

The primary free polyamines identified during growth and development of strawberry (Fragaria × ananassa Duch.) microcuttings cultivated in vitro were putrescine, spermidine and spermine. Polyamine composition differed according to tissue and stages of development; putrescine was predominant in aerial green tissues and roots. -DL-difluoromethylarginine (DFMA), a specific and irreversible inhibitor of the putrescine-synthesizing enzyme, arginine decarboxylase (ADC), strongly inhibited growth and development. Application of agmatine or putrescine to the inhibited system resulted in a reversal of inhibition, indicating that polyamines are involved in regulating the growth and development of strawberry microcuttings. -DL-difluoromethylornithine (DFMO), a specific and irreversible inhibitor of putrescine biosynthesis by ornithine decarboxylase, promoted growth and development. We propose that ADC regulates putrescine biosynthesis during microcutting development. The application of exogenous polyamines (agmatine, putrescine, spermidine) stimulated development and growth of microcuttings, suggesting that the endogenous concentrations of these polyamines can be growth limiting.Abbreviations ADC arginine decarboxylase - ODC ornithine decarboxylase - DFMA -difluoromethylarginine - DFMO -difluoromethylornithine - Put putrescine - Spd spermidine - Sp spermine - DW dry weight - PA polyamine - PPF photosynthetic photon flux     

水分胁迫下山黧豆多胺代谢与β_N_草酰_L_α,β_二氨基丙酸积累相关性的研究

Seedlings of grass pea (Lathyrus sativus L.) were treated with polyethylene glycol (PEG), in addition with putrescine (Put) and α-difluoromethylarginine (DFMA) in order to study the relation between polyamine metabolism and accumulation of β-N-oxalyl-L-α, β-diaminopropionic acid (ODAP) under water stress. The results indicated that Put, spermidine (Spd) and spermine (Spm) contents of leaves gradually increased along with the extension of PEG-treatment time. Especially, the increase in Spm contents was significant together with a gradual increase of ODAP contents. Put and Spd contents of leaves markedly increased along with the extension of PEG+Put treatment time. There was little effect on Spm contents and ODAP contents in leaves with PEG+Put treatment. The accumulation of Put, Spd, Spm and ODAP in leaves was inhibited by PEG+DFMA treatment. The inhibition of DFMA for Put and Spd biosynthesis was partially overcome by adding exogenous Put, but little effect for the inhibition of DFMA on Spm biosynthesis by adding exogenous Put. The accumulation of ODAP was inhibited with PEG+DFMA+Put treatment in leaves. This indicated that there was a relationship between the increase of Spm contents and the accumulation of ODAP in leaves under water stress.     

Involvement of polyamines in the drought resistance of rice

This study investigated whether and how polyamines (PAs) in rice (Oryza sativa L.) plants are involved in drought resistance. Six rice cultivars differing in drought resistance were used and subjected to well-watered and water-stressed treatments during their reproductive period. The activities of arginine decarboxylase, S-adenosyl-L-methionine decarboxylase, and spermidine (Spd) synthase in the leaves were significantly enhanced by water stress, in good agreement with the increase in putrescine (Put), Spd, and spermine (Spm) contents there. The increased contents of free Spd, free Spm, and insoluble-conjugated Put under water stress were significantly correlated with the yield maintenance ratio (the ratio of grain yield under water-stressed conditions to grain yield under well-watered conditions) of the cultivars. Free Put at an early stage of water stress positively, whereas at a later stage negatively, correlated with the yield maintenance ratio. No significant differences were observed in soluble-conjugated PAs and insoluble-conjugated Spd and Spm among the cultivars. Free PAs showed significant accumulation when leaf water potentials reached -0.51 MPa to -0.62 MPa for the drought-resistant cultivars and -0.70 MPa to -0.84 MPa for the drought-susceptible ones. The results suggest that rice has a large capacity to enhance PA biosynthesis in leaves in response to water stress. The role of PAs in plant defence to water stress varies with PA forms and stress stages. In adapting to drought it would be good for rice to have the physiological traits of higher levels of free Spd/free Spm and insoluble-conjugated Put, as well as early accumulation of free PAs, under water stress.     

Changes in content of free,conjugated and bound polyamines and osmotic adjustment in adaptation of vetiver grass to water deficit

Osmotic adjustment and alteration of polyamines (PAs) have been suggested to play roles in plant adaptation to water deficit/drought stress. In this study, the changes in cell intactness, photosynthesis, compatible solutes and PAs [including putrescine (Put), spermidine (Spd) and spermine (Spm) each in free, conjugated and bound forms] were investigated in leaves of vetiver grass exposed to different intensity of water deficit stress and subsequent rewatering. The results showed that, when vetiver grass was exposed to the moderate (20% and 40% PEG-6000 solutions) and severe (60% PEG solution) water deficit for 6 days, the plant injury degree (expressed as the parameters of plant growth, cell membrane integrity, water relations and photosynthesis) increased and contents of free and conjugated Put decreased with the rise of PEG concentration. Under the moderate water deficit, the plants could survive by the reduced osmotic potential (ψ_s), increased free and conjugated Spd and Spm in leaves. After subsequent rewatering, the osmotic balance was re-established, most of the above investigated physiological parameters were fully or partly recovered to the control levels. However, it was not the case for the severely-stressed and rewatering plants. It indicates that, vetiver grass can cope well with the moderate water deficit/drought stress by using the strategies of osmotic adjustment and maintenance of total contents of free, conjugated and bound PAs in leaves.     

Spermine facilitates recovery from drought but does not confer drought tolerance in transgenic rice plants expressing <Emphasis Type="Italic">Datura stramonium</Emphasis><Emphasis Type="Italic">S</Emphasis>-adenosylmethionine decarboxylase

Polyamines are known to play important roles in plant stress tolerance but it has been difficult to determine precise functions for each type of polyamine and their interrelationships. To dissect the roles of putrescine from the higher polyamines spermidine and spermine, we generated transgenic rice plants constitutively expressing a heterologous S-adenosylmethionine decarboxylase (SAMDC) gene from Datura stramonium so that spermidine and spermine levels could be investigated while maintaining a constant putrescine pool. Whereas transgenic plants expressing arginine decarboxylase (ADC) produced higher levels of putrescine, spermidine and spermine, and were protected from drought stress, transgenic plants expressing SAMDC produced normal levels of putrescine and showed drought symptoms typical of wild type plants under stress, but the transgenic plants showed a much more robust recovery on return to normal conditions (90% full recovery compared to 25% partial recovery for wild type plants). At the molecular level, both wild type and transgenic plants showed transient reductions in the levels of endogenous ADC1 and SAMDC mRNA, but only wild type plants showed a spike in putrescine levels under stress. In transgenic plants, there was no spike in putrescine but a smooth increase in spermine levels at the expense of spermidine. These results confirm and extend the threshold model for polyamine activity in drought stress, and attribute individual roles to putrescine, spermidine and spermine.