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1.
Thin cell layers excised from tobacco ( Nicotiana tabacum L. cv. Samsun) stem internodes, with an appropriate exogenous hormonal balance, were able to form a greater number of roots, and in a larger percentage of the explants (93%) than when they were excised from pedicels (40%). The developmental sequence of root formation and explant growth were followed by histological analysis. Free and bound [trichloroacetic acid (TCA)-soluble and -insoluble] putrescine and spermidine increased in the explants, particularly when root meristemoids appeared. These meristemoids originated in the superficial (day 6 in culture) or deep (days 10–11) layers and inside the newly formed callus (day 25). At those times, TCA-soluble and, to a lesser extent, TCA-insoluble bound putrescine predominated over the other polyamines. Spermine was always present in trace amounts. Polyamines decreased again when root and callus formation was completed (day 30). The involvement of these three classes of polyamines (free, TCA-soluble and -insoluble) in morphogenic processes is discussed.  相似文献   

2.
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.  相似文献   

3.
In the short-day plant, strawberry (Fragaria ananassa Duch.), polyamines (putrescine, spermidine and spermine), conjugated spermidine (water-insoluble compounds) and bound amines (putrescine, spermidine, phenylethylamine, 3-hydroxy, 4-methoxyphenylethylamine) accumulated in the shoot tips during floral induction and before floral emergence. Different associations of free amines and conjugated amines were observed during floral induction, as compared with the reproductive phase. During the whole period of floral development, phenylethylamine (an aromatic amine) was the predominant amine, representing 80 to 90% of the total free amine pool. Phenylethylamine conjugates (water-insoluble compounds) were the predominant amides observed prior to fertilization. These substances decreased drastically after fertilization. In vegetative shoot tips from plants grown continously under long days, free polyamines (putrescine, spermidine) and bound polyamines (putrescine, spermidine) were low and no change was observed. Free amines (spermine and phenylethylamine), bound aromatic amines (phenylethylamine, 3-hydroxy, 4-methoxyphenylethylamine), conjugated spermidine and phenylethylamine did not appear. Male-sterile flowers were distinguished by their lack of conjugated spermidine and phenylethyalamine and by a decrease in free phenylethylamine. In normal and sterile strawberry plants -DL-difluoromethylornithine (DFMO), a specific irreversible inhibitor of ornithine decarboxylase (ODC), caused inhibition of flowering and free and polyamine conjugates. When putrescine was added, polyamine titers and flowering were restored. A similar treatment with -DL-difluoromethylarginine (DFMA), a specific, irreversible inhibitor of arginine decarboxylase (ADC), did not affect flowering and polyamine titers. These results suggest that ornithine decarboxylase (ODC) and polyamines are involved in regulating floral initiation in strawberry. The relationship between polyamines, aromatic amines, conjugates, floral initiation and male sterility is discussed.Abbreviations ADC arginine decarboxylase - ODC ornithine decarboxylase - DFMA -DL-difluoromethylarginine - DFMO -DL-difluoromethylornithine - Put putrescine - Spd spermidine - Spm spermine - Phen phenylethylamine - 3H4M Phen 3-hydroxy, 4-methoxyphenylethylamine  相似文献   

4.
We studied the effects of various polyamines on bud regeneration in thin-layer tissue explants of vegetative and floweringNicotiana tabacum L. cv. Wisconsin 38, in which application of exogenous spermidine (Spd) to vegetative cultures causes the initiation and development of some flower buds (Kaur-Sawhney et al. 1988 Planta173, 282). We now show that this effect is dependent on the time and duration of application, Spd being required from the start of the cultures for about three weeks. Neither putrescine nor spermine is effective in the concentration range tested. Spermidine cannot replace kinetin (N6-furfurylaminopurine) in cultures at the time of floral bud formation, but once the buds are initiated in the presence of kinetin, addition of Spd to the medium greatly increases the number of floral buds that develop into normal flowers. Addition of Spd to similar cultures derived from young, non-flowering plants did not cause the appearance of floral buds but rather induced a profusion of vegetative buds. These results indicate a morphogenetic role of Spd in bud differentiation. Dedicated to Professor Hans Mohr on the occasion of his 60th birthday  相似文献   

5.
The developmental fates of shoots regenerated in culture and in situ by stem tissues of Nicotiana tabacum cv. Wisconsin 38 from different positions along the main axes of plants at different ages have been characterized. It was expected that explants from vegetative plants would not have the capacity to produce floral shoots. Contrary to the expected result, a small percentage (about 0.2%) of the shoots formed from cultured stem explants taken from young, vegetative plants were floral, i.e., produced a small number of nodes and then a flower. A larger percentage (about 2%) of the shoots formed by explants from the same region of plants which had flowered were floral. The largest percentage (76%) of floral shoots arose from explants taken from the inflorescence. Internode cells which were stimulated to divide and undergo organogenesis in situ after decapitation of the plant also produced few-noded, floral shoots with apical internode tissues producing many such floral shoots and basal internode tissues producing few such floral shoots. These results indicate that the capacity to form a flower is a visible expression of a cryptic developmental state which is quantitatively but not qualitatively controlled in time and space.  相似文献   

6.
The effects of two inhibitors of polyamine (spermidine and spermine) biosynthesis, cyclohexylamine (CHA; 5 and 10 mM) and methylglyoxal(bis-guanylhydrazone) (MGBG; 0.1, 0.5 and 1 mM), on the organogenic response in vegetative bud-forming tobacco (Nicotiana tabacum L. cv. Samsun) thin layer explants were evaluated micro- and macroscopically at different times during culture. The final number of buds formed and the percentage of organogenic explants was significantly reduced by both inhibitors, but much more so by MGBG than CHA. This inhibitory effect was already evident in MGBG-treated explants on day 5, in terms of the number of meristemoids per explant. On the contrary, in the presence of CHA, the number of meristemoids on day 5 was higher than that in the controls. Between days 9 and 13, meristemoid formation slowed down considerably in inhibitor-treated explants compared with controls. On day 13, the number of bud primordia was similar in control and CHA-treated explants, but significantly lower in MGBG-treated explants. This inhibitor also induced peculiar cytohistological events, such as a reduced formation of oval-shaped cell aggregates on the explant surface and more frequent cases of nucleolar extrusion, while CHA led to the appearance of hypertrophic epidermal cells; callus formation at the basal end of the explant and xylogenesis were also affected by the inhibitors. Ethylene biosynthesis, measured as [ C]methionine incorporation, was stimulated 2- (day 2) to 3-fold (15 h) by 0.5 mM MGBG, whereas CHA (10 mM) had little effect and aminoethoxyvinylglycine (AVG; 0.1 μM), an ethylene synthesis inhibitor, was strongly inhibitory. In control explants, the incorporation of labelled methionine into ethylene and spermidine followed an inverse trend up to day 8. In these explants, free putrescine increased 32-fold and spermidine increased about 10-fold between days 0 and 8. Trichloroacetic acid (TCA)-soluble conjugated putrescine also accumulated dramatically during culture. While CHA provoked a decline in spermidine levels, MGBG caused an unexpected increase in free spermidine and spermine titres; however, its most conspicuous effect was on the further enhancement of putrescine conjugate accumulation, while CHA and AVG had the opposite effect. Results are discussed in view of establishing a putative link between MGBG-enhanced ethylene synthesis, increased conjugate titres and inhibition of meristemoid formation.  相似文献   

7.
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  相似文献   

8.
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.  相似文献   

9.
Long-sized oligogalacturonides (OGs) are cell wall fragments that induce defence and developmental responses. The Ca2+-dependent “egg-box” conformation is required for their activity, and polyamines may prevent them from adopting this conformation. Although OGs are known to inhibit auxin-induced growth processes, their effect on cytokinin-induced ones requires investigation. In the present work OGs were shown to promote cytokinin (benzyladenine, BA)-induced vegetative shoot formation from tobacco leaf explants, independent of the presence of CaCl2 in the medium and of auxin (indoleacetic acid, IAA) supply. The effect of polyamines, putrescine (PU) and spermidine (SD) supplied with/without their biosynthetic inhibitors (DFMO, CHA) was also investigated, and showed that spermidine enhanced adventitious vegetative shoot formation, but only on medium containing Ca2+ and IAA. Treatments with inhibitors blocked this promotive effect. OGs did not alter free polyamine concentrations, but caused a moderate increase of conjugated ones, and exhibited an early inhibitory effect on polyamine biosynthetic gene expression. OGs, but not SD, caused long-term changes in calcium-associated epifluorescent signals in the cell walls, and, later, inside the cells of specific tissues. Electron microscopy analysis (ESI system) demonstrated that calcium accumulated in the cell walls and vacuoles of OG-cultured explants. The relationship between OGs, cytokinin, calcium, and polyamines in adventitious vegetative shoot formation is discussed.  相似文献   

10.
In the short-day plant Chrysanthemum (Chrysanthemum morifolium Ramat. variety Pavo) putrescine and spermidine conjugates appeared in the apical bud before the first observable transformation of the meristem into floral structures. These compounds accumulated on floral initiation and well before floral evocation. Spermidine conjugates were predominant during floral initiation whereas free amines did not accumulate to any significant extent. Different associations of amides were observed during floral initiation as compared with the reproductive phase. 3,4-Dimethoxyphenethylamine conjugates (water-insoluble compounds) were the predominant amine conjugates observed during flower development. These compounds decreased drastically after fertilization. In vegetative buds from plants grown in long days polyamine conjugates were very low and appeared as plants aged. We present evidence that ornithine decarboxylase (ODC) regulates putrescine biosynthesis during floral initiation and floral development. When ODC action was blocked by DFMO (-DL-difluoromethylornithine, a specific, irreversible inhibitor of ODC), flowering was inhibited, and free and conjugated polyamines were not detected. This treatment led to a slight enhancement of ADC activity. When putrescine was added, polyamine titers and flowering were restored. A similar treatment with DFMA (-DL difluoromethylarginine, a specific, irreversible inhibitor of ADC) did not affect flowering and the polyamine titers. The results suggest that ODC and polyamine conjugates are involved in regulating floral initiation in Chrysanthemum.Abbreviations ADC arginine decarboxylase - ODC ornithine decarboxylase - DFMA -DL-difluoromethylarginine - DFMO -DL-difluoromethylornithine  相似文献   

11.
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.  相似文献   

12.
Polyamine oxidase (PAO, EC 1.5.3.3) activity and polyamine content in the cell wall and soluble fractions obtained from embryos, endosperms and shoots and roots of etiolated or green seedlings of maize ( Zea mays L. cv. WF9) during the first 7 days of germination were investigated. Polyamine content was also determined in the trichloroacetic acid-soluble (free polyamines) and trichloroacetic acid insoluble (bound polyamines) fraction obtained from the same tissues. PAO activity, determined by the radiometric method based on the recovery of the labelled reaction product 1-pyrroline, was mostly localized in the cell wall fraction. The activity was very low in embryos and endosperms and present in traces in roots. In etiolated shoots PAO activity increased sharply, while in green shoots it was low and increased slowly. No polyamines were found in the cell wall fraction and only putrescine was detected in the soluble fraction, with the exception of the embryo, where spermidine and spermine were also present. In the TCA-soluble fraction of embryos, putrescine increased during imbibition, while spermidine and spermine decreased; in the endosperm no relevant changes in polyamines occurred. In the same fraction of green and etiolated seedlings, putrescine increased, giving a peak at days 3–5, while spermidine decreased to very low levels. The amount of bound polyamines was 1–4% of the free ones. The pattern of PAO activity seems to be unrelated to endogenous free polyamine content, which is the same in shoots and roots of etiolated and green seedlings. Enzyme activity, very low in ungerminated seeds, increased continuously during the progression of germination, especially in etiolated shoots, indicating a possible involvement in cell wall formation.  相似文献   

13.
Changes in polyamine concentration in the long day (LD) plant Rudbeckia hirta were examined over the course of floral initiation and development. Plants of R. hirta were grown to maturity under 9h, non-floral-inductive photoperiods. At maturity, half the plants were placed in ambient day length plus a 4- h night interruption. Plants were sampled at 0, 4, 8, 12 or 16 days for polyamine content and floral initiation. Polyamines were extracted from fully expanded leaves and the meristems were examined histologically. In another experiment, polyamines were extracted from the meristems under paired LD and short day (SD) conditions every 2 days from 0 to 22 days. A rise in free polyamines was linked to important cytological events during floral initiation. Free putrescine and spermidine levels increased after 4 LD and continued until 14 to 16 LD when the levels in the meristem began to decline. Events of floral initiation began between 4 and 8 LD with cell proliferation and the start of stem elongation. Initiation was irreversible after 14 to 16 LD, the period when putrescine and spermidine began to decline. After 4 LD, the polyamine level was consistently higher in the photoinduced plants. Our results from this study, using direct histological comparisons of meristematic development and polyamine concentration, clearly demonstrate a correlation of polyamines and flowering.  相似文献   

14.
Sinapis alba L. was induced to flower by either a long day or a displaced short day. Following collection of leaf (phloem) and root (xylem) exudates from induced and non-induced plants, polyamines in the exudates were extracted, separated and analyzed quantitatively. The titers of free and conjugated putrescine the major polyamine fractions in all samples, increased early and markedly in leaf exudates during the floral transition, coinciding closely with movement of the floral stimulus out of the induced leaf. By contrast, putrescine titer in the root exudate did not increase. A spray of difluoromethylornithine (DFMO), an irreversible inhibitor of the putrescine-biosynthetic enzyme ornithine decarboxylase, at hour 8 of the long day considerably reduced the titer of free and conjugated putrescine in leaf exudates, and at the same time, markedly decreased the flowering response of induced plants. This effect of DFMO on flowering was substantially reversed by a simultaneous application of putrescine to the roots. DFMO sprayed on induced plants also suppressed early activation of indices of both mitosis and DNA synthesis in the shoot apical meristem. These results support the view that the extra putrescine synthesized in induced leaves is a necessary component of the floral stimulus in Sinapis.  相似文献   

15.
离体黄瓜子叶节花芽分化与内源激素及多胺的关系   总被引:3,自引:0,他引:3  
用高效液相色谱法(HPLC)测定了黄瓜子叶节花芽分化期(0—6天)内源激素及多胺的变化。结果显示,子叶培养0—2天生长素(IAA)、赤霉素(GA3)、玉米素(ZT)、脱落酸(ABA)等4种内源激素均明显下降,4—5天略有上升,表明0-2天IAA、GA3和ABA的剧降有利于花原基形成,3—5天较高的ZT含量有利于花器官原基的形成。除腐胺(Put)外,精胺(Spm)、亚精胺(Spd)、尸胺(Cad)在0—1天均下降,1—4天上升,4—5天再下降,Put在0—1天急剧上升,而后持续下降,表明高水平的内源多胺总量和Put可能有利于花原基分化,2天后Spm含量上升有利于花器官原基分化,而Cad含量变化可能是区别花芽和营养芽分化的特征之一。  相似文献   

16.
用高效液相色谱法(HPLC)测定了黄瓜子叶节花芽分化期(0-6天)内源激素及多胺的变化。结果显示,子叶培养0-2天生长素(IAA)、赤霉素(GA_3)、玉米素(ZT)、脱落酸(ABA)等4种内源激素均明显下降,4-5天略有上升,表明0-2天IAA、GA_3和ABA的剧降有利于花原基形成,3-5天较高的ZT含量有利于花器官原基的形成。除腐胺(Put)外,精胺(Spm)、亚精胺(Spd)、尸胺(Cad)在0-1天均下降,1-4天上升,4-5天再下降,Put在0-1天急剧上升,而后持续下降,表明高水平的内源多胺总量和Put可能有利于花原基分化,2天后Spm含量上升有利于花器官原基分化,而Cad含量变化可能是区别花芽和营养芽分化的特征之一。  相似文献   

17.
Polyamines are thought to play a role in the control of inherent or environmentally-induced growth rates of plants. To test this contention, we grew plants of four grass species, the inherently fast-growing Poa annua L. and Poa trivialis L. and the inherently slow-growing Poa compressa L. and Poa pratensis (L.) Schreb., at three levels of nitrate supply. Firstly, plants were compared when grown with free access to nitrate, allowing the plants to grow at their maximum relative growth rate (RGRmax). Secondly, we compared the plants when grown with relative nitrate addition rates of 100 and 50 mmol N (mol N)–1 day–1 (RAR100 and RAR50, respectively).The freely-occurring polyamines, spermine, spermidine and putrescine, were separated from their conjugates; the latter were further subdivided into a TCA-soluble and a TCA-insoluble fraction. Each of the three fractions responded differently to the nitrate supply. Under nitrogen limitation, the total concentration of polyamines (free and bound ones together) decreased in both leaves and roots of all Poa species, whereas that in the stem remained more or less the same. These effects were to a large extent determined by the free polyamines. For the conjugates there was more differentiation, both between plant organ and among polyamine structures. A positive correlation between the RGR, LAR (leaf area per plant mass), SLA (leaf area per leaf mass), LMR (leaf mass per plant mass) and SMR (stem mass per plant mass) with the polyamine concentration was found. The RMR (root mass per plant mass) showed a negative one. No significant differences were found between the inherently fast- and slow-growing grass species.The (putrescine)/(spermine + spermidine) ratio in the leaves increased with decreasing nitrate supply, which is associated with a decrease in leaf expansion, accounting for a decrease in LAR and SLA. For the roots, this ratio tended to decrease with decreasing nitrate supply, whereas for the stems the results were somewhat more variable.We found no evidence for a crucial role of polyamines in the determination of inherent variation of growth in spite of a positive correlation of especially the free polyamines with growth parameters.  相似文献   

18.
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.  相似文献   

19.
Contents of endogenous free and conjugated polyamines were determined in embryogenic and non-embryogenic leaf regions of Camellia japonica leaf explants, before (day 0), and 20 and 45 days after the induction of direct somatic embryogenesis, to clarify whether or not polyamines are linked to the specific morphogenic responses previously reported in the leaf regions. The analysis was carried out by high-performance liquid chromatography. The results showed that there were no significant differences in the endogenous free and conjugated putrescine (Put), spermidine (Spd) and spermine (Spm) contents between the embryogenic and non-embryogenic leaf regions of the same leaf. Thus, leaf region-specific embryogenic response in C. japonica is not being determined/conditioned by the endogenous levels of Put, Spd and/or Spm. However, soluble and insoluble conjugated Put and soluble conjugated Spd seem to be related to the formation and development of globular embryos.  相似文献   

20.
The floral transition includes a complex system of factors that interact and involve various biochemical signals, including plant growth regulators. The physiological signals involved in the control of the floral transition have been sparsely studied and mainly in plant species whose genetics are poorly known. In this work, the role of polyamines, gibberellins, and cytokinins was investigated by analyzing their endogenous content in vegetative and floral buds of azalea. The results showed that there is a clear distinction between floral and vegetative buds with respect to the levels of these plant hormones, with floral buds containing higher amounts of conjugated polyamines, gibberellins (GAs) from the non-13-hydroxylation pathway (GA9, GA7, and GA4), and cytokinins (particularly isopentenyl-type species), and vegetative buds containing higher amounts of free polyamines and gibberellins from the early 13-hydroxylation pathway and fewer cytokinins. In conclusion, there is a specific pattern of endogenous hormone profiles in both vegetative and floral bud development in azalea, which may be relevant for future research on the control of flowering by exogenous hormone applications.  相似文献   

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