Polyamines promote root elongation and growth by increasing root cell division in regenerated Virginia pine (Pinus virginiana Mill.) plantlets

Polyamines have been demonstrated to play an important role in adventitious root formation and development in plants. Here, we present a detailed analysis of influence of exogenously added polyamines on adventitious root development and its relationship to cold tolerance in Virginia pine (Pinus virginia Mill.). Our results demonstrated that polyamines putrescine (Put), spermidine (Spd), and spermine (Spm) at 0.001 mM improve rooting frequency and promote root elongation. Put, Spd, and Spm at 0.01–1 mM decrease rooting frequency and reduce root elongation root elongation. Measurements of diamine oxidase (DAO, EC 1.4.3.6) and polyamine oxidase (PAO, EC 1.4.3.4) activities showed that higher DAO and PAO enzyme activities were obtained when high concentrations of polyamines were applied and when plantlets were treated for 5–7 week at 4°C and 16°C. Survival rate of plantlets increased with the treatment of polyamines at low temperature. Polyamines increased mitotic index of cells in root tips of regenerated plantlet cultured on medium containing 0.001 μM Put, Spd, or Spm, but did not increase mitotic index in tissues of needle tips of the same plantlets. These results demonstrated that polyamines promote root elongation and growth by increasing root cell division in regenerated Virginia pine plantlets.     

Polyamines reduce salt-induced oxidative damage by increasing the activities of antioxidant enzymes and decreasing lipid peroxidation in Virginia pine

Polyamines play an important role in the plant response to adverse environmental conditions including salt and osmotic stresses. In this investigation, the responses of polyamines to salt-induced oxidative stress were studied in callus cultures and plantlets in Virginia pine (Pinus virginiana Mill.). Our results demonstrated that polyamines reduce salt-induced oxidative damage by increasing the activities of antioxidant enzymes and decreasing lipid peroxidation. Among different polyamines used in this study, putrescine (Put) is more effective in increasing the activities of ascorbate peroxidase (APOX), glutathione reductase (GR), and superoxide dismutase (SOD), reducing the activities of acid phosphatase and V-type H+-ATPase, and decreasing lipid peroxidation in Virginia pine, compared to both spermidine (Spd) and spermine (Spm). When 2.1 mM Put, Spd, and Spm were separately added to the medium, higher diamine oxidase (DAO) and polyamine oxidase (PAO) activities were observed in callus cultures and plantlets, compared to the concentrations of 0.7 and 1.4 mM. The activities of these two enzymes produce hydrogen peroxide (H₂O₂), which may act in structural defense as a signal molecule and decreasing the protection of polyamines against salt-induced oxidative damage in Virginia pine.     

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.     

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.     

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.     

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 spermidine and spermine levels on salt tolerance associated with tonoplast H+-ATPase and H+-PPase activities in barley roots

The effects of polyamines (Putrescine— Put; Spermidine—Spd; and Spermine—Spm) on␣salt tolerance of seedlings of two barley (Hordeum vulgare L.) cultivars (J4, salt-tolerant; KP7, salt-sensitive) were investigated. The results showed that, the salt-tolerant cultivar J4 seedlings accumulated much higher levels of Spd and Spm and lower Put than the salt-sensitive cultivar KP7␣under salt stress. At the same time, the dry weight of KP7 decreased significantly than that of␣J4. After methylglyoxal bis(guanylhydrazone) [MGBG, an inhibitor of S-adenosylmethionine decarboxylase (SAMDC)] treatment, Spd and Spm levels together with the dry weight of both cultivars were reduced, but the salt-caused dry weight reduction in two cultivars could be reversed by the concomitant treatment with Spd. MGBG decreased the activities of tonoplast H⁺-ATPase and H⁺-PPase too, but the experiments in vitro indicated that MGBG was not able to affect the above two enzyme activities. However, the polyamines, especially Spd, promoted their activities obviously. These results suggested that the conversion of Put to Spd and Spm and maintenance of higher levels of Spd and Spm were necessary for plant salt tolerance.     

Association of polyamines in governing the chilling sensitivity of maize genotypes

Chilling stress is an important constraint of global production of maize. This study was undertaken to compare the chilling responses of different maize seedling tissues and to analyze changes in polyamines as a result of chilling stress. Reponses to chilling were characterized in two maize (Zea mays L.) inbred lines, ‘HuangC’ and ‘Mo17’, that putatively differ in chilling sensitivity. Seedlings were exposed to low temperature (5°C) and chilling injury was estimated by electrical conductivity (EC), malonaldehyde (MDA) concentration, and by changes in putrescine (Put), spermidine (Spd) and spermine (Spm) concentrations in root, mesocotyl, and coleoptile tissues. Membrane permeability (as measured by EC), MDA concentrations and Put concentrations in the three tissue of maize seedlings increased after chilling stress, except for the Put concentration in roots. Spd and Spm concentrations in the three tissues of seedlings decreased after chilling stress. The EC for cold stressed tissues were lower in HuangC than Mo17. Also, the EC of coleoptile tissues were lower than for mesocotyl in both inbred lines. We suggest that mesocotyl tissue can be used to evaluate cold tolerance in maize. Stepwise regression analyses showed that chilling injury in roots was generally correlated with Spd concentration while in the mesocotyl injury was mainly correlated with Put and Spd concentrations. Spermidine and Spm concentrations in the coleoptile were correlated with chilling injury. Characteristics changes of polyamines in chill-tolerant maize seedling combined with regression analysis are a reliable method for evaluating chill tolerance in maize lines.     

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.     

External spermine prevents UVA-induced damage of <Emphasis Type="Italic">Synechocystis</Emphasis> sp. PCC 6803 via increased catalase activity and decreased H<Subscript>2</Subscript>O<Subscript>2</Subscript> and malonaldehyde levels

Common polyamines, putrescine (Put), spermidine (Spd), and spermine (Spm), are cationic compounds known as beneficial factors for many cellular processes including cell division, proliferation, differentiation, and stress response in all living organisms. Effects of exogenous Spm on the protective responses of Synechocystis sp. PCC 6803 exposed to UVA were investigated. The presence of 0.5 mM Spm in the culture medium significantly reduced cell growth after 60 min under white light condition but protected the cells after growing for 60 min under UVA. The stress-tolerant response of Synechocystis cells represented by the ratio of putrescine/spermidine (Put/Spd) showed about a 6-fold increase after 60 min UVA in the presence of Spm. In addition, those levels of chlorophyll a, carotenoids, and photosynthetic oxygen evolution were increased by Spm supplementation in UVA-treated cells. Exogenous Spm induced the activity of catalase but not superoxide dismutase in cells under UVA treatment. On the other hand, Spm treatment enabled cells to apparently decrease the intracellular free radical H₂O₂ and malonaldehyde (MDA) levels. Overall results suggested that Spm supplementation could protect Synechocystis sp. PCC 6803 cells via the increase of Put/Spd ratio and the reduction of both H₂O₂ and MDA levels in conjunction with the induction of catalase activity. Interestingly, UVA-treated cells as compared to non-treated cells with exogenous Spm showed a decrease of Spm with an increase of Put and no change in Spd. This suggested the back conversion of Spm to Spd and finally to Put as cellular mechanism in response to UVA.     

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.     

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.     

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.     

Interactions between reactive oxygen species, ethylene and polyamines in leaves of Glycyrrhiza inflata seedlings under root osmotic stress

The interactions between reactive oxygen species (ROS), ethylene (ETH) and polyamines (PAs) in leaves of Glycyrrhiza inflata seedlings under root osmotic stress are reported. The results showed that the interactions between ROS, ETH and PAs were quite diverse at different degrees of damage. In slightly damaged leaves, the inhibition of ETH synthesis had no significant influence on ROS production and the content of putrescine (Put), spermidine (Spd) and spermine (Spm); the inhibition of Put synthesis had no significant influence on the production of ROS and ETH. However, in seriously damaged leaves, the inhibition of ETH production alleviated the increase in ROS production and the decrease in the content of Put, Spd and Spm; the reduction in polyamine content promoted the increase in the production of ROS and ETH; furthermore, exogenous H₂O₂ accelerated the increase in ETH production and the decrease in the content of these amines. Thus, it can be concluded that there is a close relationship between ROS content and the levels of ETH and PAs in the seriously damaged leaves. ROS production was modulated by the inhibition in ETH production and the reduction in polyamine content. Conversely, ROS promoted ETH production and reduced the polyamine content.     

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.     

盐胁迫下大麦幼苗多胺的种类和状态与多胺氧化酶活性的关系

200 mmol/L的NaCl胁迫8 d大麦幼苗叶片和根系中的三种形态多胺都有不同程度地下降,其中游离态多胺含量的下降幅度最大;高氯酸不溶性结合态多胺含量变化较小.根系中PAO的活性先上升后下降,而叶片中PAO的活性先下降后上升.游离态多胺中,亚精胺和精胺(Spd Spm)的含量变化与相应部位PAO的活性变化趋势相反,表明PAO在盐胁迫下可能调节了游离态多胺的含量从而影响高氯酸可溶结合态与高氯酸不溶结合态多胺的含量.     

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.     

Polyamines in the photosynthetic apparatus

The three main polyamines putrescine (Put), spermidine (Spd) and spermine (Spm) were characterized by HPLC in intact spinach leaf cells, intact chloroplasts, thylakoid membranes, Photosystem II membranes, the light-harvesting complex and the PS II complex. All contain the three polyamines in various ratios; the HPLC polyamine profiles of highly resolved PS II species (a Photosystem II core and the rection center) suggest an enrichment in the polyamine Spm.Abbreviations Chl chlorophyll - HPLC high performance liquid chromatography - LHC light-harvesting complex - PS II Photosystem II - PS II-RC Photosystem II reaction center - Put putrescine - Spd spermidine - Spm spermine - 10%S-core D₁-D₂-Cyt b559-47 kD-43 kD complex     

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.