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1.
Endogenous indole-3-acetic acid (IAA), abscisic acid (ABA) and cytokinins [zeatin (Z) zeatin riboside, dihydrozeatin, dihydrozeatin riboside, N6-isopentenyl adenine (iP) and N6-isopentenyladenine riboside] were evaluated in hazelnut (Corylus avellana L.) cotyledons of different developmental stage and genetic source for their somatic embryogenic capacity. There was an inverse correlation between the embryogenic potential of cotyledons and the degree of maturity of zygotic embryos, the first characteristic being associated with iP-type cytokinins and the second with Z-type cytokinins. Although the differences in total cytokinin, ABA and IAA contents between the cotyledons were small, the IAA/ABA and, mainly, the iP-type/Z-type cytokinin ratios were found to be two good indexes of the embryogenic competence of explants, suggesting that the endogenous hormonal balance is a very important factor defining the in vitro potential of hazelnut cotyledons. Received: 6 January 1997 / Revision received: 3 March 1997 / Accepted 1 April 1997  相似文献   

2.
Radioimmunoassays (RIA), employing antisera raised in rabbits against bovine serum albumin conjugates of zeatin riboside, dihydrozeatin riboside, and isopentenyladenosine, were used to estimate levels of these cytokinins and their corresponding bases in samples of effective (nitrogen-fixing, Fix+), ineffective (nonnitrogen-fixing, Fix) pea root nodules and uninoculated roots. Assays were done on extracts of nodule tissue, 1–2 g fresh weight, or approximately 10 g fresh weight of root tissue, and high specific activity [3H]zeatin riboside was added during preparation of the extract for use as a recovery marker. Two different purification procedures were employed, each involving several purification steps. High performance liquid chromatography (HPLC) was the final step in both procedures. Fractions from HPLC were analyzed by RIA using the appropriate antiserum. The cytokinins, zeatin, zeatin riboside, dihydrozeatin riboside, isopentenyl adenine, and isopentenyladenosine were detected and quantified in nodule tissue, and similarly, in root tissue (with the exception of zeatin, which we were unable to quantify in root tissue). Cytokinin levels in nodule tissue were higher than those in root tissue. The major cytokinins detected in nodule tissue were zeatin, followed by zeatin riboside and then dihydrozeatin riboside. The levels of zeatin and zeatin riboside estimated in nodules in the present study by RIA were of the same order of magnitude, though tending to be a little higher, than values obtained previously by bioassay. Dihydrozeatin riboside was identified with confidence for the first time in nodule tissue. There was a general decline with age in cytokinin levels in nodules, but no major qualitative change in nodule cytokinins with age. For theRhizobium strains examined, the data did not indicate a clear correlation between nodule cytokinin levels and the effectiveness of nodules in nitrogen fixation.  相似文献   

3.
Roots of young Norway spruce seedlings were incubated under hydroculture conditions in a synthetic nutrient medium containing either 3H-isopentenyladenosine, isopentenyladenosine or zeatin riboside. When feeding with 3H-isopentenyladenosine a new radiaolabelled metabolite was found in the feeding solution as well as in root extracts. Isopentenyladenosine and zeatin riboside were metabolised and for both compounds an unknown metabolite was detected in the feeding solution. The metabolites were purified by solid phase extraction, HPLC and partially characterised. A major characteristic of the metabolites is their reactivity in the presence of NH4OH, which results in the formation of the cytokinin bases isopentenyladenine or zeatin, respectively. UV-spectra and the chemical characteristics indicate that the new metabolites are closely related. The GC-MS analysis revealed, that the metabolites are true derivatives of isopentenyladenine and zeatin. The biogenesis of the new metabolites is discussed with regard to plant microbial interactions.Abbreviations Ck(s) = cytokinin(s) - GC-MS = gas chromatography-mass spectrometry - iP = isopentenyladenine - [9R]iP = isopentenyladenosine - [9G]iP = isopentenyladenine-9-glucoside - [9R-MP]iP = isopentenyladenosine-5-monophosphate - Z = trans-zeatin - [9R]Z = trans-zeatin riboside  相似文献   

4.
Hormones and Pod Development in Oilseed Rape (Brassica napus)   总被引:3,自引:0,他引:3       下载免费PDF全文
The endogenous levels of several plant growth substances (indole acetic acid, IAA; abscisic acid, ABA; zeatin, Z; zeatin riboside, [9R]Z; isopentenyladenine, iP; and isopentenyladenosine, [9R]iP were measured during pod development of field grown oilseed Rape (Brassica napus L. var oleifera cv Bienvenu) with high performance liquid chromatography and immunoenzymic (enzyme-linked immunosorbent assay, ELISA) techniques. Results show that pod development is characterized by high levels of Z and [9R]Z in 3 day old fruits and of IAA on the fourth day. During pod maturation, initially a significant increase of IAA and cytokinins was observed, followed by a progressive rise of ABA levels and a concomitant decline of IAA and cytokinin (except iP) levels. The relationship between hormone levels and development, especially pod number, seed number per pod, and seed weight determination, will be discussed.  相似文献   

5.
Development of vitrification and apical necrosis was followed in Camellia sinensis, Gerbera jamesonii, Malus domestica and hybrid Populus tremula x P. alba shoots cultured in vitro on Murashige & Skoog (MS) medium with different concentrations of growth regulators. High humidity in the culture vessels and excess of BA in the medium were found to be the major factors influencing vitrification. Lack of exogenous cytokinin in the medium during successive subcultures induced apical necrosis in poor-rooting species (Malus domestica, Camellia sinensis). The level of internal phytohormones (ABA, IAA, IPA, 2iP, Z, ZR) was determined in the apple shoots by means of ELISA. The content of internal cytokinins in the vitrified apple shoots was several times greater than in normal ones, which supports the hypothesis that excess of cytokinins, inducing rapid divisions of cells in meristems in the atmosphere with high humidity, is responsible for vitrification. Apical necrosis of the plantlets that appeared after cultivation on cytokinin-free medium is the result of deficiency in endogenous hormones in apple shoots and this being confirmed by analysis of endogenous hormones in apple shoots.Abbreviations BA benzyladenine - BHT butylated hydroxy-toluene - ABA abscisic acid - IAA indole-3-acetic acid - ELISA enzyme-linked immunosorbent assay - IPA isopentenyladenosine - 2iP isopentenyladenine - NAA naphthyl-3-acetic acid - TBS trishydroxymethylaminomethane buffered saline - TLC thin layer chromatography - Z zeatin - ZR zeatin riboside  相似文献   

6.
This paper deals with the quantitative determination of free and bound cytokinins in clubroot tissue and in Plasmodiophora brassicae Woron, infected Brassica campestris L. callus tissue. The fractions were separated in a butanol soluble fraction containing the free cytokinins such as zeatin and zeatin riboside and a water soluble fraction containing the bound cytokinins. The butanol fraction was extensively purified and analysed by high pressure liquid chromatography (HPLC). The butanol fraction contained cytokinins which cochromatographed with zeatin and zeatin riboside and not with dihydrozeatin. Zeatin and zeatin riboside were quantitatively determined by HPLC. Recovery of the cytokinins varied between 30–50%. Clubs contained 50–160 ng zeatin and 210–300 ng zeatin riboside per g dry weight. Callus tissue contained 133 ng zeatin and 169 ng zeatin riboside per g dry weight. Clubs, callus as well as healthy tissue contain large amounts of bound cytokinins. Upon treatment of the water soluble fraction first with alkaline phosphatase and then with β-glucosidase biologically active fractions were found which coeluted with zeatin and zeatin riboside on Sephadex LH20 in 20% ethanol. Evidence is presented for a novel cytokinin in the water soluble fraction which yields free zeatin and glucose-6-phosphate after treatment with β-glucosidase.  相似文献   

7.
NH4+ and K+ uptake experiments have been conducted with 3 ectomycorrhizal fungi, originating from Douglas fir (Pseudotsuga menziesii (Mirb.] Franco) stands. At concentrations up to 250 μM, uptake of both NH4+ and K+ follow Michaelis-Menten kinetics. Laccaria bicolor (Maire) P. D. Orton, Lactarius rufus (Scop.) Fr. and Lactarius hepaticus Plowr. ap. Boud. exhibit Km values for NH4+ uptake of 6, 35, and 55 μM, respectively, and Km values for K+ uptake of 24, 18, and 96 μM, respectively. Addition of 100 μM NH4+ raises the Km of K+ uptake by L. bicolor to 35 μM, while the Vmax remains unchanged. It is argued that the increase of Km is possibly caused by depolarization of the plasma membrane. It is not due to a competitive inhibition of K+ by NH4+ since the apparent inhibitor constant is much higher than the Km, for NH4+ uptake. The possibility that NH4+ and K+ are taken up by the same carrier can be excluded. The Km, values for K+ uptake in the two other fungi are not significantly affected by 100 μM NH4+. Except for a direct effect of NH4+ on influx of K+ into the cells, there may also be an indirect effect after prolonged incubation of the cells in the presence of 100 μM NH4+.  相似文献   

8.
Abstract. H+ efflux from the trap lobes of Dionaea muscipula Ellis (Venus's flytrap) was measured in vitro. FC, IAA, and 2,4-D markedly increase the rate of H+ efflux within minutes of their addition to the incubation medium whereas ABA and DES cause the rate to decrease. Consequently, the H+ efflux mechanism of Dionaea is considered to be similar to the H+ extrusion pumps of other higher plants in this respect. However, the H+ extrusion mechanism of Dionaea may be unusual in that long-term exposure of the trap lobes to known secretion elicitors— bactopeptone, NH4+, Na +, urea, thiourea, glycine or xanthine—also causes a large increase in the steady-state rate of H+ efflux from the trap lobes. Since the observed H+ effluxes primarily correspond to the adaxial surface of the trap lobes and show similar time- and secretion elicitor-dependencies to the responses seen in situ, it appears that the H+ effluxes measured in vitro bear a direct relationship to those observed in the intact, actively secreting plant. Three of the secretion elicitors that were tested— K+, NH4+, and urea—have rapid effects on the rate of H+ extrusion in addition to their long-term effects. K+ and NH4+ cause a rapid acceleration of H+ efflux whereas urea causes a rapid deceleration or, at high external concentrations, reversal of the net flux. The effect of K+ is inferred to result from K+ -H+ exchange between the tissue and bathing medium. Studies with structural analogues of NH4+ and urea and inhibitors of the assimilation of reduced nitrogen suggest that the effects of NH4+ and urea result from the pH-perturbing consequences of their metabolism subsequent to their absorption. These effects are considered to be auxiliary to the elicitation of secretion. It is proposed that H+ efflux from the trap lobes is mediated by a K+-H+ exchange mechanism, the activity of which is modified by long-term exposure to secretion elicitors and/or short-term exposure to factors which alter the availability of endogenous H+ ions.  相似文献   

9.
The production, isolation, and purification of specific chicken immunoglobulins (Igs) against three main groups of naturally occurring cytokinins are reported. The specific Igs directed against, respectively, zeatin riboside, dihydrozeatin riboside, and isopentenyladenosine are extracted from the egg yolk and used in radioimmunoassays that allow the quantification in parallel of pmol of the cytokinins in plant extracts. As little as 50 fmol of zeatin riboside, 20 fmol of isopentenyladenosine, and 40 fmol of dihydrozeatin riboside can be detected. The levels of cytokinins measured in the radio-immunoassay correlate well with physicochemical analysis methods such as high performance liquid chromatography (HPLC) with UV spectrum detection and HPLC-coupled mass spectrometric detection. Cross-reactivity studies indicate that the assay is not affected by most of the structurally related compounds. The respective antibody preparations recognized zeatin riboside, dihydrozeatin riboside, and isopentenyladenosine and the corresponding free bases. The results obtained when analyzing crude plant extracts are expressed as zeatin riboside equivalents, dihydrozeatin riboside equivalents, and isopentenyladenosine equivalents.Abbreviations B binding activity - B 0 maximal binding - B 1 unspecific binding - GC gas chromatography - HPLC high performance liquid chromatography - LC-MS HPLC-coupled mass spectrometry - MOPS 4-morpholinepropanesulfonic acid - RIA radioimmunoassay - TBS Tris-buffered saline - (diH)Z dihydrozeatin - (diH) [9R]Z dihydrozeatin riboside - iP isopentenyladenine - [9R]iP isopentenyladenosine - Z zeatin - [9R]Z zeatin riboside - [9G]iP isopentenyladenine-9-glucoside - [9R-5P]iP isopentenyladenosine-5-monophosphate  相似文献   

10.
Root nodules ofPhaseolus mungo were extracted for their cytokinin content. Four cytokininsviz. zeatin, zeatin riboside, N6(‡2-isopentenyl) aminopurine and its riboside were identified on the basis of Rf values obtained by co-chromatography with authentic samples of cytokinins on thin layer of a mixture of polyvinylpyrrolidone/CaSO4.  相似文献   

11.
Summary Four cytokinins (CKs), indole-3-acetic acid (IAA) and its ester and amino conjugates, and abscisic acid (ABA) levels of two bromeliads, Ananas comosus (L.) Merril and Vriesea gigantea Gaudich., grown in 5 mM (NH4)2SO4 or urea as the sole nitrogen (N) form, were investigated. In both bromeliads, zeatin (Z) and zeatin riboside ([9R]Z) were the most abundant CKs. In A. comosus, CKs levels decreased drastically (≊ 12 times) after 7 and 30 d in media with ammonium and urea, respectively. After 3 d in media with N, V. gigantea CK levels decreased 30 and 20 times in the presence of ammonium and urea, respectively. N-starved A. comosus and V. gigantea exhibited similar ABA levels, but ABA decreased faster in V. gigantea when plants were transferred to media with N. Free IAA levels decreased until the 15th and 30th day when A. comosus was transferred to a medium with ammonium and urea, respectively. N-starved A. comosus amide, ester, and free IAA amounted to 81%, 14%, and 4%, respectively. There was a transient increase in the proportion of amide IAA and a corresponding decrease of the ester and the free IAA proportion when N-starved plants were transferred to media with N. The relationship between the internal hormonal patterns and the different ecological adaptations of the two bromeliads are discussed.  相似文献   

12.
Mixed nitrate/ammonium fertilization can partially alleviate the negative effects of salinity on growth of some plant species compared to all-nitrate or all-ammonium fertilization. To gain insights about the mechanisms involved, tomato (Solanum lycopersicum L. cv Moneymaker) plants were grown hydroponically for 3 weeks with two NO3 /NH4 + fertilization regimes (6/0.5 and 5/1.5; Ntotal = 6.5 mM) in the absence (control) or presence of salt stress (100 mM NaCl). Ammonium enrichment had no effect on growth and other parameters under control conditions. Under salinity, however, ammonium enrichment improved shoot and root biomass by 20% and maintained leaf PSII efficiency close to control levels. These changes were related to higher leaf K+, NO3 , and NH4 + concentrations and activities of the N-assimilatory enzymes glutamate synthase (GOGAT) and glutamine synthase (GS) in the leaves. Ammonium enrichment also attenuated the salt-induced increase in leaf abscisic acid (ABA) concentration and decrease in leaf concentrations of indole 3-acetic acid (IAA) and the cytokinins trans-zeatin (tZ) and trans-zeatin riboside (tZR). Enhanced cytokinin status was probably due to maintenance of root-to-shoot cytokinin transport and decreased leaf induction of the cytokinin-degrading enzyme cytokinin oxidase/dehydrogenase (CKX) under ammonium-enriched conditions. It is concluded that nitrogen form modifies salinity-induced physiological responses and that these modifications are associated with changes in plant hormone status.  相似文献   

13.
Tomato growth was examined in solution culture under constant pH and low levels of NH4+ or NO3?. There were five nitrogen treatments: 20 mmoles m?3 NH4+, 50 mmoles m?3 NO3?, 100 mmoles m?3 NH4+ 200 mmoles m?3 NO3?, and 20 mmoles m?3 NH4++ 50 mmoles m?3 NO3?. The lower concentrations (20 mmoles m?3 NH4+ and 50 mmoles m?3 NO3?) were near the apparent Km for net NH4+ and NO3? uptake; the higher concentrations (100 mmoles m?3 NH4+ and 200 mmoles m?3 NO3?) were near levels at which the net uptake of NH4+ or NO3? saturate. Although organic nitrogen contents for the higher NO3? and the NH4++ NO3? treatments were 22.2–30.3% greater than those for the lower NO3? treatment, relative growth rates were initially only 10–15% faster. After 24 d, relative growth rates were similar among those treatments. These results indicate that growth may be only slightly nitrogen limited when NH4+ or NO3? concentrations are held constant over the root surface at near the apparent Km concentration. Relative growth rates for the two NH4+ treatments were much higher than have been previously reported for tomatoes growing with NH4+ as the sole nitrogen source. Initial growth rates under NH4+ nutrition did not differ significantly (P≥ 0.05) from those under NO3? or under combined NH4++ NO3?. Growth rates slowed after 10–15 d for the NH4+ treatments, whereas they remained more constant for the NO3? and mixed NH4++ NO3? treatments over the entire observation period of 24–33 d. The decline in growth rate under NH4+ nutrition may have resulted from a reduction in Ca2+, K+, and/or Mg2+ absorption.  相似文献   

14.
In M. braunii, the uptake of NO3 and NO2 is blue-light-dependent and is associated with alkalinization of the medium. In unbuffered cell suspensions irradiated with red light under a CO2-free atmosphere, the pH started to rise 10s after the exposure to blue light. When the cellular NO3 and NO2 reductases were active, the pH increased to values of around 10, since the NH4+ generated was released to the medium. When the blue light was switched off, the pH stopped increasing within 60 to 90s and remained unchanged under background red illumination. Titration with H2SO4 of NO3 or NO2 uptake and reduction showed that two protons were consumed for every one NH4+ released. The uptake of Cl was also triggered by blue light with a similar 10 s time response. However, the Cl -dependent alkalinization ceased after about 3 min of blue light irradiation. When the blue light was turned off, the pH immediately (15 to 30 s) started to decline to the pre-adjusted value, indicating that the protons (and presumably the Cl) taken up by the cells were released to the medium. When the cells lacked NO3 and NO2 reductases, the shape of the alkalinization traces in the presence of NO3 and NO2 was similar to that in the presence of Cl, suggesting that NO3 or NO2 was also released to the medium. Both the NO3 and Cl-dependent rates of alkalinization were independent of mono- and divalent cations.  相似文献   

15.
High performance liquid chromatography analysis of immunoaffinity-purified extracts of mycelia of Amanita muscaria, and the Amaranthus bioassay of the eluted fractions, revealed the following seven cytokinins: zeatin, zeatin riboside, zeatin N-9-glucoside, dihydrozeatin, dihydrozeatin riboside, isopentenyl adenine, and isopentenyl adenosine. The decreased growth of aluminum-treated mycelia correlated with a 35% decrease in the total amount of the cytokinins. Among individual cytokinins, zeatin was the most affected, exhibiting a reduction of about 90%. The results are compared with previous investigations of aluminum effects on cytokinins in the mycelia of Lactarius piperatus, whose growth is stimulated by aluminum.Abbreviations ZR zeatin riboside - iPA isopentenyl adenosine - Z zeatin - DHZ dihydrozeatin - iP isopentenyl adenine - DHZR dihydrozeatin riboside - Z-9G zeatin N-9-glucoside - iP-9G isopentenyl N-9-glucoside - HPLC high performance liquid chromatography - DHZRMP dihydrozeatin riboside monophosphate - ZRMP zeatin riboside monophosphate  相似文献   

16.
Effects of cytokinins were studied on rotenone-sensitive NADH dehydrogenase in mitochondria from fresh potato tubers (Solarium tuberosum), in consideration of the operation of external and rotenone-insensitive internal NADH dehydrogenases that has not been fully accounted for in previous studies. In submitochondrial particles (smp), zeatin was only weakly active, and zeatin riboside (ZR) was inactive. Inhibition rates at 400 μM of isopentenyladenine (iP) and isopentenyladenosine (iPA) were 45% and 30%, respectively, and that of BA (BA) was 64%. In intact mitochondria, the inhibition by iP and BA significantly increased, I50 being 50 and 250 μM, respectively, but that by zeatin and iPA decreased. A structure–activity study showed that hydrophobic and steric factors are important for the activity. Cytokinins inhibited the electron flow via natural quinone more strongly than that via synthetic quinone. These results suggest that among the cytokinins the species that can regulate the electron transport is iP rather than its riboside or zeatin.  相似文献   

17.
Potato cells grown in liquid culture incorporated mevalonic acid lactone-[2-14C] into free cytokinin (zeatin riboside and zeatin and the cytokinin of RNA (zeatin riboside). The cytokinin liberated by catabolism of RNA can account for no more than 40% of the free cytokinins.  相似文献   

18.
The naturally occurring cytokinins, zeatin, zeatin riboside and dihydrozeatin did not promote the germination of celery (Apium graveolens L.) seeds and 6-Δ2-isopentenyladenine (2iPA) and its riboside were only moderately active. Of the synthetic cytokinins, kinetin, kinetin riboside, and the disubstituted urea, N-phenyl-N′-pyridyl urea (NC5392) were moderately active, and 6-benzyl-aminopurine (BA) and its derivatives BA riboside and 6-benzyl-amino-9(tetrahydropyran-2yl)purine (SD8339) were the most active cytokinins tested. 6-(o-hydroxybenzyl)aminopurine (hyd-BA) and its naturally occurring riboside inhibited germination under normally inductive conditions. All the cytokinins examined were more active in promoting germination of lettuce (Lactuca sativa L.) than celery seeds. BA, BA riboside and SD8339 were again the most active cytokinins. In contrast to the results with celery, zeatin and zeatin riboside were highly active. The other cytokinins also showed high activity with the exception of dihydrozeatin, hyd-BA and hyd-BA riboside which were less active. Cytokinin ribosides were less active than the corresponding free bases during the early period of the lettuce seed incubation but total germination after 90 h was similar.  相似文献   

19.
Ian M. Scott  Roger Horgan 《Planta》1984,161(4):345-354
the cytokinins of tobacco crown-gall tissue have been analysed by quantitative mass spectrometry using 2H2-labelled cytokinin riboside 5-monophosphates and 15N4-labelled cytokinin glycosides as internal standards. The principal endogenous cytokinin of this tissue is zeatin riboside 5-monophosphate. The biologically inactive 7-glucoside of zeatin is the most abundant basic cytokinin in the tissue. These findings expose the limitations of previously reported analyses of similar tissues, which were restricted to biologically active basic cytokinins. The present study demonstrates that the endogenous cytokinins of tobacco crowngall tissue show a clear correspondence to the range of metabolites formed when exogenous cytokinins are supplied to nontumorous tobacco cells.Abbreviations DHZ dihydrozeatin - DHZ7G dihydrozeatin 7-glucoside - DHZMP dihydrozeatin 9-riboside 5-monophosphate - DHZR dihydrozeatin 9-riboside - GC-MS coupled gas chromatography-mass spectrometry - HPLC high-performance liquid chromatography - Z7G zeatin 7-glucoside - Z9G zeatin 9-glucoside - ZOG zeatin O-glucoside - ZMP zeatin 9-riboside 5-monophosphate - ZR zeatin 9-riboside - ZROG zeatin 9-riboside O-glucoside  相似文献   

20.
In an attempt to optimize somatic embryo formation in Oncidium ‘Gower Ramsey’, the effects of five auxins (2,4-D, IAA, IBA, NAA and picloram) and five cytokinins (2iP, BA, kinetin, TDZ and zeatin), used alone, was tested in vitro using root-derived callus. In general, kinetin (0.5 and 2 mg l−1) and zeatin (0.5 mg l−1) were found to be more effective than other auxin and cytokinin treatments to induce somatic embryogenesis from root-derived callus.  相似文献   

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