Changes in endogenous polyamines during flower development in two diverse species of rose

Changes in the concentrations of endogenous free, conjugated and bound polyamine were determined in petals of two different species of rose, viz. Rosa damascena and Rosa bourboniana, from small bud (stage 1) till full bloom (stage 8). High free putrescine and spermidine concentrations were associated with early stages of flower development and then decreased in R. damascena. At full bloom, the concentration of free putrescine was higher than rest of the polyamines measured. A steady increase in conjugated putrescine, spermidine and spermine was observed during entire period of flower development with predominance of conjugated putrescine at full bloom. In R. damascena the bound spermine was higher than rest of the polyamines during full bloom. In R. bourboniana, during the early stages of flower development, similar situation was observed, however, at full bloom, free spermidine concentration was higher than rest of the polyamines. In this species, the concentration of conjugated and bound spermine was higher than rest of the polyamines during full bloom. Polyamine concentrations were generally lower in the petals of R. bourboniana than R. damascena which may be due to genotypic differences. The possible roles of the observed polyamines are discussed in relation to flower development.IHBT Communication no, 0345.     

Changes in free polyamine titers and expression of polyamine biosynthetic genes during growth of peach <Emphasis Type="Italic">in vitro</Emphasis> callus

In the present paper, correlation between free polyamines and growth of peach (Prunus persica cv. Yuzora) in vitro callus was investigated. Growth of the callus was divided into three phases based on measurement of fresh weight. Free polyamines, putrescine (Put), spermidine (Spd), and spermine (Spm), could be detected during peach callus growth. Changes in free Put titers followed the callus growth rate, as shown by low and stable levels in the first stage, quick increase at the beginning of the second phase, and slow increase in the last phase, whereas fluctuations of Spd and Spm titers were aberrant from that of Put at early stage. Expressions of five key genes involved in polyamine biosynthesis were characterized, in which only the genes leading to Put synthesis, ADC (arginine decarboxylase) and ODC (ornithine decarboxylase), agreed with callus growth and fluctuation of Put titers. Treatment of the callus with D-arginine, an inhibitor of ADC, led to significant growth inhibition and enormous reduction of endogenous Put, coupled with obvious decrease of mRNA levels of ADC and ODC. Exogenous application of Put partially restored the callus growth, along with resumption of endogenous Put and expression levels of ADC and ODC. Spd and Spm titers experienced minor change in comparison to Put. The data presented here suggested that free Put played an important part in peach callus growth. Putative mechanisms or mode of action underlying the role of Put in peach callus growth and different expression patterns of the genes responsible for polyamine biosynthesis are also discussed.     

Tissue-specific changes in polyamine levels of neural tissue and fat body in adult crickets

The three major polyamines—putrescine, spermidine, and spermine—were studied and changes of their levels were examined in extracts of cerebral ganglia and fat body from adult Acheta domesticus. In nervous tissue, only spermidine and spermine were present and spermine was two- to three-fold more abundant than spermidine. The polyamine levels were high up to day 3, decreased on day 4, and then remained relatively unchanged up to day 10. The spermidine/spermine ratios decreased during the imaginal life. Higher spermidine titres were observed in the neural tissue of egg-laying females compared to virgin females. In the fat body, putrescine was detected together with spermidine and spermine. Spermidine and spermine levels were two-fold higher than putrescine. Fat body of virgin females contained two times more polyamines than male fat body. Low at emergence, spermidine and spermine concentrations peaked on days 2–3 only in females, and egg-laying was characterized by an increase of putrescine and spermidine titres. Starvation did not change polyamine contents, implying homeostatic regulation of the intracellular polyamine metabolism. These data showing tissue specific changes in polyamine levels during the imaginal life of Acheta domesticus point to the physiological importance of polyamines as possible intracellular regulators during adult insect development. © 1993 Wiley-Liss, Inc.     

Polyamine changes during the development of zygotic embryos in Prunus avium

A study of the polyamine profile was carried out during zygotic embryo development in Prunus avium. Zygotic embryos were collected from 2 donor trees and sorted into 3 size classes: C1 [2.5 to 3.5 mm]; C2 [3.6 to 4.5 mm] and C3 [5.5 to 7 mm]. Evolution of the various polyamines was similar for the two donor trees. Changes in the relative amount of the various free polyamines were observed during zygotic embryo development. Agmatine and spermine levels increased from C1 to C3. Spermidine, the predominant polyamine, showed a two-fold decrease in C3 compared with C1 and C2; the evolution of putrescine was opposed, showing an increase in the last developmental stage. The putrescine/spermidine ratio could be a marker for these 3 developmental stages with a higher ratio in C3 compared with C1 and C2. Polyamine changes in cotyledons from class C1 were investigated during in vitro culture. A 10-day induction on a medium containing 2,4-dichlorophenoxyacetic acid (2,4-D) and kinetin caused a strong decline in free spermidine levels and a dramatic increase in free putrescine. The formation of conjugated putrescine occurred simultaneously, and twenty days after removal of growth regulators, the various polyamine contents were still at the same level.Abbreviations Agm agmatine - Dap diaminopropane - 2,4-D 2,4-dichlorophenoxyacetic acid - Put putrescine - Spd spermidine - Spm spermine     

Comparison of soybean cultivars for enhancement of the polyamine contents in the fermented soybean natto using Bacillus subtilis (natto)

Polyamines have beneficial properties to prevent aging-associated diseases. Raw soybean has relatively high polyamine contents; and the fermented soybean natto is a good source of polyamines. However, detailed information of diversity of polyamine content in raw soybean is lacking. The objectives of this study were to evaluate differences of polyamines among raw soybeans and select the high polyamine-containing cultivar for natto production. Polyamine contents were measured chromatographically in 16 samples of soybean, which showed high variation among soybeans as follows: 93–861 nmol/g putrescine, 1055–2306 nmol/g spermidine, and 177–578 nmol/g spermine. We then confirmed the high correlations of polyamine contents between raw soybean and natto (r = 0.96, 0.95, and 0.94 for putrescine, spermidine, and spermine, respectively). Furthermore, comparison of the polyamine contents among 9 Japanese cultivars showed that ‘Nakasen-nari’ has the highest polyamine contents, suggesting its suitability for enhancement of polyamine contents of natto.     

Recombinant polyamine-binding protein of <Emphasis Type="Italic">Synechocystis</Emphasis> sp. PCC 6803 specifically binds to and is induced by polyamines

His-tagged Synechocystis sp. PCC 6803 PotD protein (rPotD) involved in polyamine transport was overexpressed in Escherichia coli. The purified rPotD showed saturable binding kinetics with radioactively labeled polyamines. The rPotD exhibited a similar binding characteristic for three polyamines, with putrescine having less preference. The K _d values for putrescine, spermine, and spermidine were 13.2, 8.3, and 7.8 μM, respectively. Binding of rPotD with polyamines was maximal at pH 8.0. Docking of these polyamines into the homology model of Synechocystis PotD showed that all three polyamines are able to interact with Synechocystis PotD. The binding modes of the docked putrescine and spermidine in Synechocystis are similar to those of PotF and PotD in E. coli, respectively. Competition experiments showed specific binding of rPotD with polyamines. The presence of putrescine and spermidine in the growth medium could induce an increase in PotD contents, suggesting the role of PotD in mediating the transport of polyamine in Synechocystis sp. PCC 6803.     

Changes in free polyamines in Tulipa gesneriana flower stalks during dry-storage and after planting of bulbs

Tulip bulbs cv. Apeldoorn are dry-stored at 5°C for 12 weeks to ensure sufficient elongation of the flower stalk, when subsequently planted at higher temperatures (17–20°C). To investigate whether free polyamines are involved in this process, flower stalk internodes were analyzed during dry-storage and after planting of the bulbs.During dry-storage for 12 weeks at 5°C (cooled) and 17°C (non-cooled), the free putrescine, spermidine and spermine amounts per flower stalk increased. The putrescine amount increased at 5°C significantly more than at 17°C, whereas the opposite was found for the spermine amount. These differences developed early during dry-storage and disappeared rapidly at subsequent higher temperatures.After planting, the lower- and uppermost flower stalk internodes of the pre-cooled bulbs elongated much faster than those of the non-cooled ones. In the pre-cooled bulbs, the free polyamine amounts per internode increased with time after planting, but the time course of these changes was different. In the non-cooled bulbs, the free polyamine amounts increased to a much lesser extent or remained more or less constant.It is argued that the observed changes in the free polyamine contents are probably not required for the cold-induced extension growth of tulips cv. Apeldoorn.Abbreviations PA polyamine - Put putrescine - Spd spermidine - Spm spermine     

Cadaverine: a common polyamine in zygotic embryos and somatic embryos of the species Capsicum chinense Jacq.

The behavior of endogenous polyamines was studied in somatic embryos and zygotic embryos of Habanero pepper (Capsicum chinense). In the first part of the work, the polyamine content was evaluated in both types of embryos (somatic and zygotic). As a result, in addition to the common polyamines (putrescine, spermidine and spermine), it was also possible to detect cadaverine, a polyamine rarely found in plants. In general, all the polyamines were found to be more abundant in somatic embryos than in zygotic embryos, with significantly higher contents of putrescine and cadaverine. Subsequently, the content of putrescine, spermidine, spermine and cadaverine, in their different forms (free, bound and conjugated) was determined in somatic embryos which were cultured in non-ventilated and ventilated containers. Detection of polyamines was carried out at 28 and 42 days of culture by the HPLC method. The ethylene content was monitored during the process in both culture conditions (ventilated and non-ventilated). As a result of the analysis, cadaverine was always found present, indicating that it is a common polyamine in the species. Ethylene was detected in containers without ventilation throughout the culture, except during replenishment of the culture medium (R1, R2 and R3). The behavior pattern of each polyamine, analyzed under different culture conditions (ventilated and non-ventilated) and at different moments of culture (28 and 42 days of culture), show that the polyamines are not only involved in morphogenic processes in plants; polyamines are also significantly affected by the surrounding environment. However, the most novel result, presented for the first time in this paper, is that cadaverine is found to be a common polyamine in C. chinense since it is present in both zygotic embryos and somatic embryos.

     

The role of free polyamines in the alternate-bearing of pistachio (<Emphasis Type="Italic">Pistacia vera</Emphasis> cv. Pontikis)

The present study was conducted in order to examine the physiological role of free polyamines in flower bud abscission. For this reason five 15-year old pistachio trees cv. "Pontikis" were selected and half of the main branches were manually defruited in early May. Polyamines were analyzed in three different organs (shoots, leaves and flower buds) from both fruiting and non-fruiting branches, during the period of kernel growing. Five samplings took place and the polyamines putrescine, spermidine and spermine were assayed. The flower bud abscission percentage was recorded every 5–10 days during kernel formation. Polyamine concentration declined during the period that coincides with that of kernel development, in both fruiting and non-fruiting branches, while significant bud abscission occurred from mid-July till late September in fruiting branches. Polyamine concentration in organs from fruiting branches was in most cases lower than that of non-fruiting ones. Most of the individual polyamines exhibited a high and significant negative correlation with bud abscission. By measuring the spermidine content of leaves and the spermine content of buds, it was possible to estimate the forthcoming bud abscission with significant accuracy (approximately 93%). On the other hand, the total polyamine content of the buds exhibited a significant strong negative relationship with bud abscission. Consequently, polyamines could have an important physiological function in the development of flower bud abscission of pistachio.     

Polyamine biosynthesis and titer during various developmental stages of Phaseolus vulgaris

The activities of arginine decarboxylase (ADC; EC 4.1.1.19) and ornithine decarboxylase (ODC; EC 4.1.1.17) as well as polyamine content were examined in Phaseolus vulgaris L. cv. Taylor's Horticultural before and during anthesis, during fruit development and throughout vegetative growth. The specific activities of polyamine biosynthetic enzymes were highest in all rapidly growing tissues, e.g., root apices, hypocotyls, young internodes, young leaves, flower buds, young pods and pericarps. They were lowest in mature, non-growing tissues. Similarly, the content of the major polyamines (putrescine, spermidine, spermine) is highest in rapidly growing tissues, and lowest in mature tissue. These correlations reinforce the growing connection between polyamines and rates of cell division and metabolic activity during both vegetative and reproductive development.     

伏令夏橙愈伤组织体细胞胚发生中多胺水平的变化

以继代培养8年的伏令夏橙愈伤组织为材料,研究了不同类型愈伤组织体细胞胚发生能力的差异和多胺水平的变化及两者之间的关系.结果表明,胚性愈伤组织的多胺含量高于非胚性愈伤组织,体细胞胚发生能力与多胺水平呈正相关.体细胞胚发生早期Put含量的增加有利于体细胞胚发生.球形胚大量形成时,Spd达到最高值;球形胚发育后期并有少量心形胚形成时Spm达到峰值.随着倍性的增加,伏令夏橙体细胞胚发生能力降低.精氨酸脱羧酶的活性变化与Put水平呈正相关,表明它是调节伏令夏橙体细胞胚发生中多胺水平的重要因子.     

Profile of polyamines during sprouting and growth of saffron (Crocus sativus L.) Corms

The profile of free and conjugated polyamines putrescine, spermidine, and spermine was studied at the onset of sprouting and during various stages of vegetative growth in saffron (Crocus sativus L.) corms. Polyamines were extracted from the shoot meristems and estimated by high performance liquid chromatography. Free putrescine was not detected at the onset of sprouting, whereas free spermidine and spermine levels increased rapidly on sprouting and decreased during further stages of corm development. The levels of conjugated polyamines were several times higher than the free forms indicating their possible role in the developmental processes. A comparison of polyamine levels of vegetative and floral corms showed higher titers of free polyamines in vegetative and conjugated polyamines in floral corms.     

Biochemical changes during seed development in <Emphasis Type="Italic">Pinus taeda</Emphasis> L.

Amino acids, polyamines, 3-indoleacetic acid (IAA), abscisic acid (ABA), buffer-soluble protein and starch contents and dry matter accumulation were analyzed in megagametophytes containing developing embryos during seed development in Pinus taeda. The highest total amino acids and polyamine contents occurred at the cotyledonary stage, followed by a significant decrease in the mature seed. Free polyamines exhibited higher levels than conjugated ones, with putrescine being the predominant type until the cotyledonary stage, and spermidine at the mature seed stage. IAA content increased continually from the globular stage reaching the maximum at the cotyledonary stage, followed by a decrease in the mature seed. The highest ABA level occurred at the globular stage, followed by a continuous reduction until stabilization at the pre-cotyledonary stage. Buffer-soluble protein and starch contents, and dry matter increased progressively during development, reaching their maximum values at the mature stage.     

Fluctuation in free and conjugated polyamines in Scots pine seedlings after changes in temperature and daylength

Free, soluble and insoluble conjugated polyamines from the needles, roots and stem of five month old Scots pine (Pinus sylvestris L.) seedlings inoculated with Suillus variegatus (Fr.) O. Kuntze and seedlings without inoculation were analysed during decrease in daylength and temperature. Temporary changes in free, soluble and insoluble conjugated polyamine pools caused by a decrease in daylength or temperature were observed. Inoculation of pine seedlings affected significantly the polyamine levels of five month old pine seedlings. The roots of inoculated seedlings contained significantly higher levels of free and soluble conjugated purtrescine and free, soluble conjugated and insoluble conjugated spermidine than the roots of noninoculated seedlings. The needles of inoculated seedlings contained significatly higher concentrations of free putrescine and soluble conjugated spermidine but lower amount of free spermine than the needles of noninoculated seedlings. The stems of inoculated seedlings contained higher concentrations of free putrescine but lower amounts of insoluble conjugated spermine. Changes in polyamine levels in noninoculated seedlings were observed after shortening of the daylength, whereas in inoculated ones changes were induced mainly by the decrease in temperature. The possible role of polyamines in the initial stage of cold hardening process is discussed.     

Polyamines and their biosynthetic enzymes during somatic embryo development in red spruce (<Emphasis Type="Italic">Picea rubens</Emphasis> Sarg.)

Summary The major objective of this study was to determine if the observed changes in polyamines and their biosynthetic enzymes during somatic embryo development were specifically related to either the stage of the embryo development or to the duration of time spent on the maturation medium. Somatic embryos of red spruce (Picea rubens) at different developmental stages, grown in the embryo development and maturation media for various lengths of time, were separated from the associated subtending tissue (embryogenic and the suspensor cell masses) and analyzed for their polyamine content as well as for polyamine biosynthetic enzyme activities. Polyamine content was also analyzed in embryos representing different stages of developmentthat were collected from the sam culture plate at the same time and the subtending tissue surrouding them. Putrescine was the predominant polyamine in the pro-embryogenic tissue, while spermidine was predominant during embryo development. Significant changes in spermidine/putrescine and spermine/putrescine ratios were observed at all stages of embryo development as compared to the pro-embryogenic cell mass. Changes in the ratios of various polyamines were clearly correlated with the developmental stage of the embryo rather than the period of growth in the maturation medium. Whereas the activities of both ornithine decarboxylase and arginine decarboxylase increased by week 3 or 4 and stayed high during the subsequent 6 wk of growth, the activity of S-adenosylmethionine decarboxylase steadily declined during embryo development.     

Ornithine decarboxylase transgene in tobacco affects polyamines,in vitro-morphogenesis and response to salt stress

Transgenic tobacco plants expressing the putrescine synthesis gene ornithine decarboxylase from mouse were raised to study the effects of up-regulation of a metabolic pathway as critical as the polyamine biosynthesis on the plant growth and development, in vitro-morphogenesis and their response to salt stress. Further, the response of the alternate pathway (arginine decarboxylase) for putrescine synthesis to the modulation of the ornithine decarboxylase pathway has also been investigated. The over-expression of the odc gene and increased levels of putrescine in tobacco led to a delay in plant regeneration on selection medium which could be overcome by the exogenous application of polyamine biosynthesis inhibitors and spermidine. Further, the lines generated had a variable in vitro morphogenic potential, which could be correlated to the shifts in their polyamine metabolism. These studies have brought forward the critical role played by polyamines in the normal development of plants and also their role in plant regeneration. Since polyamines are known to accumulate in cells under abiotic stress conditions, the tolerance of the transgenics to salt stress was also investigated and the transgenics with their polyamine metabolism up-graded showed increased tolerance to salt stress.     

Inhibition of the ethylene response by 1-MCP in tomato suggests that polyamines are not involved in delaying ripening, but may moderate the rate of ripening or over-ripening

Ethylene initiates the ripening and senescence of climacteric fruit, whereas polyamines have been considered as senescence inhibitors. Ethylene and polyamine biosynthetic pathways share S-adenosylmethionine as a common intermediate. The effects of 1-methylcyclopropene (1-MCP), an inhibitor of ethylene perception, on ethylene and polyamine metabolism and associated gene expression was investigated during ripening of the model climacteric fruit, tomato (Solanum lycopersicum L.), to determine whether its effect could be via polyamines as well as through a direct effect on ethylene. 1-MCP delayed ripening for 8 d compared with control fruit, similarly delaying ethylene production and the expression of 1-aminocyclopropane-1-carboxylic acid (ACC)-synthase and some ethylene receptor genes, but not that of ACC oxidase. The expression of ethylene receptor genes returned as ripening was reinitiated. Free putrescine contents remained low while ripening was inhibited by 1-MCP, but increased when the fruit started to ripen; bound putrescine contents were lower. The activity of the putrescine biosynthetic enzyme, arginine decarboxylase, was higher in 1-MCP-treated fruit. Activity of S-adenosylmethionine-decarboxylase peaked at the same time as putrescine levels in control and treated fruit. Gene expression for arginine decarboxylase peaked early in non-treated fruit and coincident with the delayed peak in putrescine in treated fruit. A coincident peak in the gene expression for arginase, S-adenosylmethionine-decarboxylase, and spermidine and spermine synthases was also seen in treated fruit. No effect of treatment on ornithine decarboxylase activity was detected. Polyamines are thus not directly associated with a delay in tomato fruit ripening, but may prolong the fully-ripe stage before the fruit tissues undergo senescence.     

水稻胚与胚乳分化发育中的内源多胺

稻胚发育过程中,其内源多胺以腐胺、亚精胺为主。在幼胚分化期,腐胺和亚精胺的含量很高;幼胚分化完成时,其含量急剧下降;直至分化后期才趋稳定。在胚及胚乳发育时期,还出现一种未知多胺X_(22),其含量除在胚分化完成时较少外,在胚发育的其他各期中,含量则一直很高。DNA和蛋白质含量的变化,从分化期开始递增直至物质积累成熟期,其趋势均相同。多胺可能参与胚与胚乳中核酸和蛋白质合成的调节。     

Endogenous polyamine content during in vivo maturation and in vitro culture of maize pollen

Changes in polyamine content during in vivo maturation and in vitro culture of maize (Zea mays L.) pollen were studied. The endogenous content of free, conjugated and bound polyamines was analyzed during 30 days of pollen evolution, in both developmental pathways (microsporogenesis and androgenesis). The induction of androgenesis from cold-pretreated uninucleate pollen results, in most of cases, in a lower total polyamine content than that of the in vivo uninucleate pollen. These differences indicate that polyamine metabolism is altered during the induction of androgenesis, and this could be a consequence of increased polyamine assimilation. In general, pollen stages that involve cell division (tetrades, pre-anthesis pollen and four-day cultured pollen) are characterized by a predominance of free Spd. The increase of Spd and Spm in 15-day cultured pollen, when the first embryoids are formed, outline the possible implication of these polyamines in embryogenetic processes. Furthermore, these findings may contribute to the improvement of maize androgenesis yield, especially in recalcitrant genotypes, by the exogenous application of polyamines or polyamine-inhibitors to the culture medium.Abbreviations PAs polyamines - Put putrescine - Spd spermidine - Spm spermine - S free polyamine fraction - SH conjugated polyamine fraction - PH bound polyamine fraction     

Polyamine homeostasis in transgenic plants overexpressing ornithine decarboxylase includes ornithine limitation

It was reported recently that overexpression of human ornithine decarboxylase (ODC) cDNA in transgenic rice plants resulted in increased steady-state concentration of polyamines, i.e., enough biosynthetic control is invested at this step to enable adjustment of polyamine levels. To investigate critically whether constitutive overexpression of ODC is sufficient to control steady-state polyamine levels, we expressed an ODC cDNA from Datura stramonium in transgenic tobacco plants. Transgenic progeny of self-fertilised primary transformants exhibited increases in ODC activity of 25-fold in leaves and 5-fold in flower buds. However, the increase in putrescine levels was only 1.5- to 2.1-fold in leaves and 1.1- to 1.3-fold in flower buds. Emphatically, no changes to spermidine or spermine steady-state levels or to soluble or insoluble hydroxycinnamic acid-conjugated polyamines were observed. Ornithine feeding to cell suspension cultures derived from the transgenic plants indicated that putrescine accumulation was limited in part by ornithine availability. These results demonstrate that a large increase in the capacity of the tobacco plants to decarboxylate ornithine does not result in a comparable increase in the level of free or conjugated polyamines. Plant polyamine homeostatic mechanisms efficiently accommodate increased ODC activity, suggesting that polyamine biosynthetic control is invested at multiple interdependent steps.