Distribution of polyamines and their related catabolic enzyme in etiolated and light-grown leguminosae seedlings

Diamine-oxidase (DAO; EC 1.4.3.6) activity and di-and polyamine levels were estimated along the epicotyl and root of light-grown and etiolated lentil (Lens culinaris Medicus) and pea (Pisum sativum L.) seedlings. The activity of DAO was higher in etiolated epicotyls than in lightgrown ones. In both species there was a positive correlation between DAO activity and the diamine (putrescine and cadaverine) levels along the whole epicotyl and root. Polyamine (spermine and spermidine) distribution seemed to be associated with the meristematic and elongating zone of the epicotyl and root. The physiological function of DAO is discussed in relation to its possible role in providing hydrogen peroxide to peroxidase-dependent reactions occurring in the cell wall.Abbreviations CAD cadaverine - DA diamine - DAO diamine oxidase - PA polyamine - PUT putrescine - SPD spermidine - SPM spermine     

Amine levels in Lathyrus sativus seedlings during development

In growing Lathyrus sativus seedlings, the levels of DNA, RNA and protein markedly decreased in the cotyledons and progressively increased in the embryo-axis. In cotyledons, spermidine and spermine contents were substantially reduced while those of agmatine and putrescine were sharply increased. By contrast the embryo-axis progressively accumulated relatively larger amounts of agmatine, homoagmatine. putrescine, cadaverine, spermidine and spermine in parallel with similar changes in its DNA, RNA and protein content. While the cotyledons contained ca 50% of the total agmatine and putrescine present in the plant embryo by day 10, the embryo-axis, though representing less than 20% of the dry wt, contained 90 and 75% of total cadaverine and homoagmatine respectively of the seedlings. Spermidine and spermine levels of this tissue were also comparatively higher, being of the order of 80 and 50% respectively of the total. The root and shoot portions of the embryo-axis also exhibited a similar relationship between changes in DNA, RNA and protein and all the above amines during development. However, the polyamine content of the shoots was relatively higher than those of the roots during the growth period.     

Distribution of cadaverine and other amines in higher plants

The distribution of the diamines cadaverine and putrescine and of the polyamines spermidine and spermine has been determined in pea seedlings at variou     

Herbicide-Induced Diamine Accumulation in Pea Roots. The Effect of Napropamide on Polyamine Levels

Pea (Pisum sativum L. cv. Alaska) root tips were incubated innutrient medium for 24 h prior to herbicide treatment. Endogenouslevels of putrescine, cadaverine, spermidine and spermine weredetermined in root tip sections at various times following 10and 20µM napropamide application. Little effect on polyaminelevels occurred within 12 h of treatment. However, by 48 h oftreatment, putrescine levels increased more than 3-fold in eachof the root sections within 4 mM of the tip while cadaverineincreased by 2-fold in only the root section 4 to 10 mM fromthe tip. After 72 h of herbicide treatment, the level of putrescineincreased over 8-fold within 5 mM of the root tip. Napropamidehad little effect on spermidine and spermine concentration within10 mM of the root tip after 72 h of exposure. This representsthe first report of an herbicideinduced accumulation of putrescineand cadaverine and suggests a possible phytotoxic effect ofhigh concentrations of putrescine on roots. (Received December 8, 1987; Accepted September 20, 1988)     

苹果幼苗侧根形成与内源多胺关系的通径分析(英文)

本文采用偏回归和通径分析研究苹果幼苗侧根形成与内源多胺含量之间的关系。多胺对苹果幼苗侧根形成有重要的调节作用,内源多胺的水平高低与侧根数量之间有显著的线性相关。腐胺、亚精胺和精胺对侧根形成的影响不同,它们对侧根数量的直接通径系数分别是0.0756,0.1558和0.8023。因而表明精胺在增加侧根数量中的作用最大,而亚精胺和腐胺几乎没有直接的作用。不过,亚精胶和腐胺通过影响精胺水平,对侧根形成有很大的间接作用。     

Changes in Polyamine Titer in Etiolated Pea Seedlings Following Red Light Treatment

The polyamines agmatine, cadaverine, putrescine, spermidineand spermine were measured by means of thin layer chromatographyand high performance liquid chromatography in buds and in 5mm long subapical sections of the 3rd internode of 6-day-oldetiolated pea seedlings. The polyamine pattern of each organwas specific, relative quantities varying with age and growth.While agmatine, putrescine, spermidine and spermine were presentin buds and in tissues of the 3rd internode, cadaverine wasfound in the 3rd internode only. Concentrations of spermidineand spermine were higher in the bud than in the 3rd internode,and the highest putrescine titer was found in the internode.Short exposure of etiolated seedlings to red light (5 min) increasedbud development while inhibiting growth of the 3rd internode.In general, exposure to red light increased the titer of putrescine,agmatine and spermidine in the bud, whereas in the internodea reverse pattern was found, i.e., internodes of seedlings growingin the dark yielded higher titer of polyamines in general, andagmatine in particular. These results are particularly pronounced18 hr after exposure to red light. A link between phytochrome-controlledgrowth and polyamine titer is suggested. ² On sabbatical leave from the Hebrew University of Jerusalem,Department of Horticulture, Rehovot, Israel. ³ Supported by a grant from the Turkish Government; Permanentaddress: Department of General Botany, University of Istanbul,Suleymaniye, Istanbul, Turkey. ¹ Supported by a grant from NSF to A.W.G. (Received August 24, 1981; Accepted October 22, 1981)     

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.     

Influence of diamines and polyamines on the senescence of plant suspension cultures

The diamines putrescine and cadaverine and the polyamines spermine and spermidine inhibited the senescence of nonphotosynthetic cultures of Paul's Scarlet rose. Response was observed when the media of stationary phase cultures was adjusted to either 1 mM of cadaverine or putrescine; or 0.1 μM of either spermine or spermidine along with 2% sucrose in all cases. Senescence of the cultures was followed by microscopic examination of cell aliquots removed at 10 day intervals and treated with the vital stain, fluorescein diacetate.     

Polyamines protect against DNA strand breaks and aid cell survival against irradiation in Escherichia coli

Polyamines protected plasmid DNA strand breaks in vitro and aided the cell survival against irradiation in polyamine-deficient Escherichia coli mutant strain. DNA strand breaks were prevented 4–6 fold more by spermidine and spermine than by putrescine and cadaverine in the dithiothreitol/Fe(III)/O2 system. After UV-irradiation, the protection of DNA strand breaks by spermine and spermidine was twice as effective as that by putrescine and cadaverine. Survivability of polyamine-deficient Escherichia coli mutant cells grown in the medium containing putrescine and spermidine was 2.4- and 3.0-fold as high as in polyamine-depleted medium at a dose of 60 and 40 J/m2. After -irradiation to a dose of 80 Gy, cell survivals of a mutant strain were significantly increased to 7.7- and 23.8-fold by putrescine and spermidine, respectively. These results implicate the possibility that polyamines play a potent role in the protection of DNA or cell damage by radiation. © Rapid Science Ltd. 1998     

Occurrence of cadaverine in hairy roots of Brugmansia candida

The polyamine, cadaverine, was detected in transformed root cultures of Brugmansia candida (syn. Datura candida), a Solanaceae which produces the tropane alkaloids scopolamine and hyoscyamine. To the best of our knowledge, this is the first time that the existence of this uncommon polyamine has been detected in a Datura species. Cadaverine, however, could not be found in the whole plant. The occurrence of cadaverine in hairy roots could be a consequence of either the transformation or a response to stress. Also, cadaverine could be participating in other secondary pathways rather than to the tropane alkaloids. The common polyamines, putrescine, spermidine and spermine were also observed.     

Polyamine-stimulated phosphorylation of proteins from corn (Zea mays L.) coleoptiles

The effect of polyamines (spermine, spermidine and putrescine) on in vitro phosphorylation of proteins from corn coleoptiles was investigated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography. Spermine promoted the phosphorylation of several membrane and soluble proteins and most of the proteins phosphorylated were different from those phosphorylated in the presence of calcium. Spermidine promoted the phosphorylation to a lesser extent and putrescine had very little stimulatory effect. Spermine-promoted phosphorylation of soluble proteins was dependent upon the presence of Mg2+ and was discernible at 100 microM spermine concentration.     

Synthesis and content of polyamines in bloodstream Trypanosma brucei

The sensitive dansyl procedure was used to detect putrescine and spermidine, but not spermine and cadaverine, in pleomorphic Trypanosoma brucei. The polyamines were synthesized in vitro from [3H]ornithine, [14C]arginine and [14C]methionine. Proline, agmatine, and citrulline, but not glutamine, glutamic or pyroglutamic acids, stimulated spermidine formation from [4C]methionine. Putrescine and sperimidine synthesis occurred rapidly from ornithine: putrescine synthesis peaked in 0.5 h, spermidine in 1 h. Trypanosoma brucei assimilated exogenous 14C-labeled putrescine, spermidine, and spermine; spermidine and spermine were taken up 5 times as rapidly as putrescine. Polyamine syntheses may therefore be a practical target for novel trypanocies.     

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.     

Retarded growth of an Escherichia coli mutant deficient in spermidine synthase can be unspecifically repaired by addition of various polyamines

The Escherichia coli mutant speE deficient in the gene encoding for spermidine synthase has no absolute requirement for spermidine but shows a retarded growth rate. This growth retardation could be unspecifically restored to the respective wild type level by exogenously supplied polyamines such as spermidine, spermine and homospermidine as well as the diamines putrescine and cadaverine. In comparison to the respective wild type, the mutant shows a two-fold increased level of endogenous putrescine but displays a reduced ability to accumulate the diamines putrescine and cadaverine. The ability to accumulate polyamines is not affected. The deleted spermidine synthase gene of the mutant was substituted by heterologous expression of the hss gene from Rhodopseudomonas viridis encoding homospermidine synthase.     

Changes in polyamine pattern are involved in floral initiation and development in Polianthes tuberosa

In the day-neutral plant Polianthes tuberosa (cv. Double) putrescine and spermine in corms at the early floral initiation stage decreased by 26 and 36%, respectively, compared with that in the vegetative stage. In contrast, a sharp increase in spermidine and cadaverine titers in corms was recorded at the early floral initiation stage. However, cadaverine in corms disappeared at the flower development stage. Polyamines in the roots were generally lower than those in the leaves and corms. In no case was the change in endogenous polyamine titers in the roots and leaves associated with floral initiation and flower development in P. tuberosa. Exogenous application of spermidine at 5, 25 or 150 microg per plant at the vegetative stage did not affect flower primordium counts. However, addition of a spermidine synthase inhibitor, cyclohexylamine, at 150 or 250 microg per plant (each dose was applied two times in total at an interval of 4 days) significantly reduced flower primordium counts, indicating that spermidine is involved in floral initiation and floral development in P. tuberosa. In P. tuberosa corms at the vegetative stage arginine decarboxylase activity rises and decreases at the early floral initiation stage. In contrast, ornithine decarboxylase activity reaches the highest level at the early floral initiation stage and declines significantly at the vegetative stage. Results indicate that an increase in spermidine and a transient increase in cadaverine titers in the corms seem characteristic of early floral initiation in P. tuberosa. It is also suggested that a significant reduction in putrescine and spermine in the corms is involved in the early floral initiation in P. tuberosa.     

Hydroxyl radical scavenging and singlet oxygen quenching properties of polyamines

Polyamines (cadaverine, putrescine, spermidine, spermine) have been shown to be present in all prokaryotic and eukaryotic cells, and proposed to be important anti-inflammatory agents. Some polyamines at high concentrations are known to scavenge superoxide radicals in vitro. We have investigated the possible antioxidant properties of polyamines and found that polyamines, e.g., cadaverine, putrescine, spermidine and spermine do not scavenge superoxide radicals at 0.5, 1.0 and 2 mM concentrations. However, polyamines were found to be potent scavengers of hydroxyl radicals. Hydroxyl radicals were produced in a Fenton type reaction and detected as DMPO-OH adducts by electron paramagnetic resonance spectroscopic technique. Spermine, spermidine, putrescine and cadaverine inhibited DMPO-OH adduct formation in a dose dependent manner, and at 1.5 mM concentration virtually eliminated the adduct formation. The *OH-dependent TBA reactive product of deoxyribose was also inhibited by polyamines in a dose-dependent manner. Polyamines were also found to inhibit the 1O2-dependent 2,2,6,6-tetramethylpiperidine N-oxy 1 (TEMPO) formation. 1O2 was produced in a photosensitizing system using Rose Bengal or Methylene Blue as photosensitizers, and was detected as TEMP-1O2 adduct by EPR spectroscopy. Spermine or spermidine inhibited the 1O2-dependent TEMPO formation maximally to 50%, whereas putrescine or cadaverine inhibited this reaction only up to 15%, when used at 0.5 and 1 mM concentrations. These results suggest that polyamines are powerful. OH scavengers, and spermine or spermidine also can quench singlet oxygen at higher concentrations.     

A possible role of cadaverine in the biosynthesis of polyamines in the Japanese newt testis

Polyamine content in testes of various vertebrates was studied extensively. Putrescine, spermidine and spermine were detected in all the animals examined, although the distribution pattern varied greatly from animal to animal. Cadaverine was detected only in amphibian testes; sym-homospermidine was found not only in testes but also in various other tissues of amphibians and of some reptiles. In the newt testis the concentration of cadaverine was lower than that of any other polyamines in summer, but there was a great increase in cadaverine content from autumn to winter. The testicular content of cadaverine was greater than that of other polyamines in winter. There was a gradual decrease in the cadaverine content in spring. The spermidine and spermine levels, which were rather low in winter, increased in spring and reached a peak in summer when spermatogenesis was active. The testicular concentration of putrescine that was much higher than that of spermidine or spermine throughout the year, increased only a little in summer. There was a significant negative correlation between the cadaverine levels and four other polyamine levels. Exogenous cadaverine decreased the testicular levels of putrescine. Mammalian gonadotropins decreased the cadaverine levels and increased the levels of other polyamines. A partially purified LH fraction from pituitaries of bullfrog, Rana catesbeiana, was also potent in depleting cadaverine of the testes of newts kept at 8 degrees C. These results suggest that testicular cadaverine suppresses the biosynthesis of polyamines, especially spermidine and spermine which are closely associated with spermatogenesis.     

Involvement of Polyamines with Adventitious Root Development in Stem Cuttings of Mung Bean

Spermine enhances the number of adventitious roots developingon stem cuttings of Phaseolus aureus Roxb. This effect is observedwhen spermine is supplied alone to cuttings or in the presenceof indolebutyric acid (IBA). That concentration most effectivein inducing the rooting response also enhances root growth.Other concentrations tested were without effect on growth. Spermidinedoes not influence root number or growth except at high concentration,when it is inhibitory to number only. Methylglyoxal bis(guanylhydrazone)(MGBG) inhibits rooting and root growth in the presence or absenceof IBA. Treatment of stem-cuttings with IBA leads to enhancedlevels of spermine, spermidine and putrescine in the hypocotylprior to development of any root primordia. MGBG reduces thelevels of spermine and spermidine whilst increasing the levelof putrescine. Furthermore, MGBG prevents the IBA-induced increasein spermine and markedly inhibits that in spermidine. Theseresults are consistent with an essential role for polyaminesand their metabolism in the early events which lead to adventitiousroot development. (Received January 10, 1983; Accepted March 17, 1983)     

Binding sites of the polyamines putrescine, cadaverine, spermidine and spermine on A- and B-DNA located by simulated annealing

Molecular dynamics simulations with simulated annealing are performed on polyamine-DNA systems in order to determine the binding sites of putrescine, cadaverine, spermidine and spermine on A- and B-DNA. The simulations either contain no additional counterions or sufficient Na+ ions, together with the charge on the polyamine, to provide 73% neutralisation of the charges on the DNA phosphates. The stabilisation energies of the complexes indicate that all four polyamines should stabilise A-DNA in preference to B-DNA, which is in agreement with experiment in the case of spermine and spermidine, but not in the case of putrescine or cadaverine. The major groove is the preferred binding site on A-DNA of all the polyamines. Putrescine and cadaverine tend to bind to the sugar-phosphate backbone of B-DNA, whereas spermidine and spermine occupy more varied sites, including binding along the backbone and bridging both the major and minor grooves.