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1.
The compartmentation and metabolism of indole-3-acetic acid (IAA) was examined in protoplasts derived from needles ofPinus sylvestris L., leaves of normal plants ofNicotiana tabacum L., leaves ofN. tabacum plants carrying the T-DNA gene 1 (rG1 plants) and leaves ofN. tabacum plants carrying the T-DNA gene 2 (rG2 plants) by using a rapid cell-fractionation method. In all tissues, 30%–40% of the IAA pool was located in the chloroplast,
while the remainder was found in the cytosol. Quantitative analysis of indole-3-ethanol (IEt) showed that in bothPinus andNicotiana the IEt pool was located exclusively in the cytosol. The only plant that contained endogenous indoleacetamide (IAAm) was
therG1-mutant ofN. tabacum, expressing theAgrobacterium tumefaciens T-DNA gene 1. Cellular fractionation of protoplasts from this transgenic plant showed that the entire IAAm pool was located
in the cytosol. Feeding experiments utilizing [5-3H]tryptophan, [5-3H]IEt, [1′-14C] and [2′-14C]IAA demonstrated that the biosynthesis and catabolism of IAA occurred in the cytosol in bothPinus and in the wild type and the different mutants ofNicotiana. Furthermore, the biosynthesis of IAAm in therG1 plants was also shown to be localized in the cytosol. 相似文献
2.
Abstract The catabolism of indole-3-acetic acid was investigated in chloroplast preparations and a crude enzyme fraction derived from chloroplasts of Pisum sativum seedlings. Data obtained with both systems indicate that indole-3-acetic acid undergoes decarboxylative oxidation in pea chloroplast preparations. An enhanced rate of decarboxylation of [1′-1C]indole-3-acetic acid was obtained when chloroplast preparations were incubated in the light rather than in darkness. Results from control experiments discounted the possibility of this being due to light-induced breakdown of indole-3-acetic acid. High performance liquid chromatography analysis of [2′-14C]indole-3-acetic acid-fed incubates showed that indole-3-methanol was the major catabolite in both the chloroplast and the crude enzyme preparations. The identification of this reaction product was confirmed by gas chromatography-mass spectrometry when [2H5]indole-3-methanol was detected in a purified extract derived from the incubation of an enzyme preparation with 32H5]indole-3-acetic acid. 相似文献
3.
Gas chromatography-mass spectrometric analyses of purified extracts from cultures of Rhizobium phaseoli wild-type strain 8002, grown in a non-tryptophan-supplemented liquid medium, demonstrated the presence of indole-3-acetic acid (IAA), indole-3-ethanol (IEt), indole-3-aldehyde and indole-3-methanol (IM). In metabolism studies with 3H-, 14C- and 2H-labelled substrates the bacterium was shown to convert tryptophan to IEt, IAA and IM; IEt to IAA and IM; and IAA to IM. Indole-3-acetamide (IAAm) could not be detected as either an endogenous constituent or a metabolite of [3H]tryptophan nor did cultures convert [14C]IAAm to IAA. Biosynthesis of IAA in R. phaseoli, thus, involves a different pathway from that operating in Pseudomonas savastanio and Agrobacterium tumefaciens-induced crown-gall tumours.Abbreviations IAA
indole-3-acetic acid
- IAld
indole-3-aldehyde
- IAAm
indole-3-acetamide
- IEt
indole-3-ethanol
- IM
indole-3-methanol
- HPLC-RC
high-performance liquid chromatography-radio counting
- GC-MS
gas chromatography-mass spectrometry 相似文献
4.
Göran Sandberg 《Planta》1984,161(5):398-403
Combined gas chromatography-mass spectrometry has been used to identify indole-3-ethanol (IEt) in a purified extract from needles of Pinus sylvestris L. Quantitative estimates obtained by high-performance liquid chromatography with fluorescence detection, corrected for samples losses occurring during purification, indicate that Pinus needles contain 46±4 ng g-1 IEt. This compares with 24.5±6.5 ng g-1 indole-3-acetic acid (IAA) and 2.3±0.4 ng g-1 indole-3-carboxylic acid (ICA) (Sandberg et al. 1984, Phytochemistry, 23, 99–102). Metabolism studies with needles incubated in a culture medium in darkness revealed that both [3-14C]-tryptophan and [2-14C]tryptamine mine are converted to [14C]IEt. It was also shown that [3-14C]IEt acted as a precursor of [14C]IAA. The observed metabolism appears to be enzymic in nature. The [2-14C]IAA was not catabolised to [14C]ICA in detectable quantities implying that, at best, only a minor portion of the endogenous ICA pool in the Pinus needles originates from IAA.Abbreviations DEAE
diethylaminoethyl
- GC-MS
gas chromatography-mass spectrometry
- HPLC
high-performance liquid chromatography
- IAA
indole-3-acetic acid
- ICA
indole-3-carboxylic acid
- IEt
indole-3-ethanol
- PVP
polyvinylpyrrolidone 相似文献
5.
Summary High perfomance liquid chromatography (HPLC) of the products of [5-3H] tryptophan metabolism byFrankia sp. Avc I1 indicates that small amounts of [3H] indole-3-acetic acid (IAA) are excreted into the growth medium.Frankia has a limited capacity for the catabolism of [2-14C]IAA and the product that accumulates is different from that detected inRhizobium japonicum cultures following inoculation with [2-14C]IAA. The data imply that the rate of turnover of IAA is much more rapid inRhizobium thanFrankia and that the two organisms employ different routes for the catabolism of IAA. 相似文献
6.
J. Sánchez-Bravo A. Ortuño J. M. Botía J. A. Del Río M. Caballero M. Acosta F. Sabater 《Plant Growth Regulation》1990,9(4):315-327
The products of indole-3-acetic acid (IAA) metabolism by incubating hypocotyl sections and decapitated seedlings of Lupinus albus were investigated. Single treatments using [1-14C]-IAA, [2-14C]-IAA or [5-3H]-IAA and double treatments using [1-14C]-IAA+[5-3H]-IAA were carried out. Extracts from treated plant material were analyzed by paper chromatography (PC), Thin layer chromatography (TLC), and high performance liquid chromatography (HPLC). When hypocotyl sections were incubated in [2-14C]-IAA, several IAA decarboxylation products including indole-3-aldehyde (IA1), indole-3-methanol (IM), 3-hydroxymethyloxindole (HMOx), methyleneoxindole (MOx) and 3,3-bisindolylmethane (BIM) were detected in the 95% ethanol extract; a latter extraction with 1M NaOH rendered IAA, IM and BIM, suggesting that conjugated auxins were formed in addition to conjugated IM. In sections incubated with [1-14C]-IAA, the 1M NaOH extraction also produced IAA so confirming the formation of conjugated auxins. The same decarboxylation products and two conjugated auxins, indole-3-acetylaspartic acid (IAAsp) and 1-O-(indole-3-acetyl)--D-glucose (IAGlu), were detected in the acetonitrile extracts from decapitated seedlings treated with [5-3H]-IAA. After a double isotope treatment ([1-14C]-IAA+[5-3H]-IAA) of decapitated seedlings, the ratio 14C/3H measured in the HPLC fractions of the acetonitrile extracts confirmed the presence of decarboxylation products as well as conjugated auxins. 相似文献
7.
Germinating seed ofDalbergia dolichopetala converted both [2H5]l-tryptophan and [2H5]indole-3-ethanol to [2H5]indole-3-acetic acid (IAA). Metabolism of [2-14C]IAA resulted in the production of indole-3-acetylaspartic acid (IAAsp), as well as several unidentified components, referred to as metabolites I, II, IV and V. Re-application of [14C]IAAsp to the germinating seed led to the accumulation of the polar, water-soluble compound, metabolite V, as the major metabolite, together with a small amount of IAA. Metabolites I, II and IV were not detected, nor were these compounds associated with the metabolism of [2-14C]IAA by shoots and excised cotyledons and roots from 26-d-oldD. dolichopetala seedlings. Both shoots and cotyledons converted IAA to IAAsp and metabolite V, while IAAsp was the only metabolite detected in extracts from excised roots. The available evidence indicates that inDalbergia, and other species, IAAsp may not act as a storage product that can be hydrolysed to provide the plant with a ready supply of IAA.Abbreviations HPLC-RC
high-performance liquid chromatography-radiocounting
- IAA
indole-3-acetic acid
- IAAsp
indole-3-acetylaspartic acid
- IAlnos
2-O-indole-3-acetyl-myo-inositol
- IEt
indole-3-ethanol 相似文献
8.
The tryptophan auxotroph mutant trp3-1 of Arabidopsis thaliana (L.) Heynh., despite having reduced levels of l-tryptophan, accumulates the tryptophan-derived glucosinolate, glucobrassicin and, thus, does not appear to be tryptophan-limited.
However, due to the block in tryptophan synthase, the mutant hyperaccumulates the precursor indole-3-glycerophosphate (up
to 10 mg per g FW). Instability of indole-3-glycerophosphate leads to release of indole-3-acetic acid (IAA) from this metabolite
during standard workup of samples for determination of conjugated IAA. The apparent increase in “conjugated IAA” in trp3-1 mutant plants can be traced back entirely to indole-3-glycerophosphate degradation. Thus, the levels of neither free IAA
nor conjugated IAA increase detectably in the trp3-1 mutant compared to wild-type plants. Precursor-feeding experiments to shoots of sterile-grown wild-type plants using [2H]5-l-tryptophan have shown incorporation of label from this precursor into indole-3-acetonitrile and indole-3-acetic acid with
very little isotope dilution. It is concluded that Arabidopsis thaliana shoots synthesize IAA from l-tryptophan and that the non-tryptophan pathway is probably an artifact.
Received: 1 March 2000 / Accepted: 10 April 2000 相似文献
9.
Mary Helen M. Goldsmith 《Planta》1982,155(1):68-75
The velocity of transport and shape of a pulse of radioactive indole-3-acetic acid (IAA) applied to a section of maize (Zea mays L.) coleoptile depends strongly on the concentration of nonradioactive auxin in which the section has been incubated before, during, and after the radioactive pulse. A pulse of [3H]IAA disperses slowly in sections incubated in buffer (pH 6) alone; but when 0.5–5 M IAA is included, the pulse achieves its maximum velocity of about 2 cm h-1. At still higher IAA concentrations in the medium, a transition occurs from a discrete, downwardly migrating pulse to a slowly advancing profile. Specificity of IAA in the latter effect is indicated by the observation that benzoic acid, which is taken up to an even greater extent than IAA, does not inhibit movement of [3H]IAA. These results fully substantiate the hypothesis that auxin transport consists of a saturable flux of auxin anions (A-) in parallel with a nonsaturable flux of undissociated IAA (HA), with both fluxes operating down their respective concentration gradients. When the anion site saturates, the movement of [3H]IAA is nonpolar and dominated by the diffusion of HA. Saturating polar transport also results in greater cellular accumulation of auxin, indicating that the same site mediates the cellular efflux of A-. The transport inhibitors napthylphthalamic acid and 2,3,5-triiodobenzoic acid specifically block the polar A- component of auxin transport without affecting the nonsaturable component. The transport can be saturated at any point during its passage through the section, indicating that the carriers are distributed throughout the tissue, most likely in the plasmalemma of each cell.Abbreviations A-
auxin anion
- HA
undissociated auxin
- IAA
indole-3-acetic acid
- NPA
N-1-napthylphthalamic acid
- TIBA
2,3,5-triiodobenzoic acid 相似文献
10.
Binding of fusicoccin (FC) to microsomal preparations of corn (Zea mays L.) coleoptiles is enhanced after incubation of the tissue with indole-3-acetic acid (IAA). Treatment of the kinetic data according to Scatchard shows that the enhancement is a consequence of an increase in the number of high-affinity FC-binding sites without changes of their KD. The minimal effective concentration of IAA is 10-7 M; above 10-5 M the effect declines. The stimulation is insensitive to protein-synthesis inhibitors (cycloheximide and puromycin). The same effect is observed with the synthetic auxins 2,4-dichlorophenoxyacetic acid and naphtalene-1-acetic acid while it is abolished by the auxin antagonists naphtalene-2-acetic acid and p-chlorophenoxyisobutyric acid. Since the above effect is only observed with intact tissue and not after incubation of IAA with microsomal preparations, a direct interaction of IAA with the FC-binding sites is ruled out and an alternative mechanism must be sought.Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- FC
fusicoccin
- [3H]FC
3H-labeled dihydrofusicoccin
- IAA
indole-3-acetic acid
- 1-NAA
naphtalene-1-acetic acid
- 2-NAA
naphtalene-2-acetic acid
- PCIB
p-chlorophenoxyisobutyric acid 相似文献
11.
12.
Translocation of Radiolabeled Indole-3-Acetic Acid and Indole-3-Acetyl-myo-Inositol from Kernel to Shoot of Zea mays L. 总被引:1,自引:1,他引:0
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Either 5-[3H]indole-3-acetic acid (IAA) or 5-[3H]indole-3-acetyl-myo-inositol was applied to the endosperm of kernels of dark-grown Zea mays seedlings. The distribution of total radioactivity, radiolabeled indole-3-acetic acid, and radiolabeled ester conjugated indole-3-acetic acid, in the shoots was then determined. Differences were found in the distribution and chemical form of the radiolabeled indole-3-acetic acid in the shoot depending upon whether 5-[3H]indole-3-acetic acid or 5-[3H]indole-3-acetyl-myo-inositol was applied to the endosperm. We demonstrated that indole-3-acetyl-myo-inositol applied to the endosperm provides both free and ester conjugated indole-3-acetic acid to the mesocotyl and coleoptile. Free indole-3-acetic acid applied to the endosperm supplies some of the indole-3-acetic acid in the mesocotyl but essentially no indole-3-acetic acid to the coleoptile or primary leaves. It is concluded that free IAA from the endosperm is not a source of IAA for the coleoptile. Neither radioactive indole-3-acetyl-myo-inositol nor IAA accumulates in the tip of the coleoptile or the mesocotyl node and thus these studies do not explain how the coleoptile tip controls the amount of IAA in the shoot. 相似文献
13.
Oxindole-3-acetic acid (OxIAA) has been identified in germinating seeds of Scots pine (Pinus sylvestris) using gas chromatography-mass spectrometry. Seeds germinated for 5 d contained 2.7 ng OxIAA·g-1 (dry weight) whereas ungerminated seeds contained 0.2 ng·g-1. Isotopically labelled OxIAA was formed in seeds incubated with [1-14C]-, [2-14C]- or [2H5]indole-3-acetic acid.Abbreviations DDC
sodium diethyldithiocarbamate
- GC
gas chromatography
- HPLC
high-performance liquid chromatography
- IAA
indole-3-acetic acid
- MS
mass spectrometry
- OxIAA
oxindole-3-acetic acid
- PVP
polyvinylpyrrolidone
- TMS
trimethylsilyl 相似文献
14.
By means of gas chromatography-selected ion monitoring-mass spectrometry using an isotope-dilution assay with 4,5,6,7-tetradeutero-indole-3-acetic acid as the internal standard, indole-3-acetic acid has been estimated to be present in aseptically cultured gametophytes of wild-type Physcomitrella patens (Hedw.) B.S.G. at a level of 0.075 g g–1 dry weight or 2.1 ng g–1 fresh weight.Abbreviations IAA
indole-3-acetic acid
- d4IAA
4,5,6,7-tetra-deutero-indole-3-acetic acid
- [14C]IAA
indole-3-[2-14C]-acetic acid
- GC-SIM-MS
gas chromatography-selected ion monitoring-mass spectrometry 相似文献
15.
Synthesis of indole-3-acetic acid (IAA), using stable-isotope incorporation, was investigated in Zea mays L. Incorporation of 2H from 2H2O into IAA molecules was shown to occur in intact plantlets and excised primary roots cultured in vitro. This demonstrates the de-novo formation of IAA, a process which is quantitatively well defined and is initiated early in germination.Abbreviations IAA
indole-3-acetic acid 相似文献
16.
Jean Rigaud 《Archives of microbiology》1970,72(4):297-307
Summary Among the indole compounds formed when tryptophan 2-14C is metabolized by Rhizobium, indole-3-lactic acid (ILA) is specially studied. In the course of experiments carried out in the culture medium of growing Rhizobium and in suspensions of washed bacterial cells the amount of ILA formed is compared with that of indole-3-acetic acid (IAA) occurring simulataneously. The formation of ILA and that of IAA directly depend on a transamination reaction. A large quantity of ILA is present in suspensions of washed bacterial cells.When ILA alone, as precursor, is incubated with Rhizobium, several products are identified: IAA, indole-3-acetaldehyde and tryptophol. Tryptophan is also detected in the aqueous fraction and is labelled when ILA 2-14C is used. The pathway of this metabolism are discussed and a general scheme is suggested. 相似文献
17.
The promoter of the nit1 gene, encoding the predominantly expressed isoform of the Arabidopsis thaliana (L.) Heynh. nitrilase isoenzyme family, fused to the β-glucuronidase gene (uidA) drives β-glucuronidase expression in the root system of transgenic A. thaliana and tobacco plants. This expression pattern was shown to be controlled developmentally, suggesting that the early differentiation
zone of root tips and the tissue surrounding the zone of lateral root primordia formation may constitute sites of auxin biosynthesis
in plants. The root system of A. thaliana was shown to express functional nitrilase enzyme. When sterile roots were fed [2H]5-L-tryptophan, they converted this precusor to [2H]5-indole-3-acetonitrile and [2H]5-indole-3-acetic acid. This latter metabolite was further metabolized into base-labile conjugates which were the predominant
form of [2H]5-indole-3-acetic acid extracted from roots. When [1-13C]-indole-3-acetonitrile was fed to sterile roots, it was converted to [1-13C]-indole-3-acetic acid which was further converted to conjugates. The results prove that the A. thaliana root system is an autonomous site of indole-3-acetic acid biosynthesis from L-tryptophan.
Received: 3 February 1998 / Accepted: 17 April 1998 相似文献
18.
Developing lime fruit [Citrus aurantifolia (Christm.) Swingle] was supplied with dl-tryptophan-3-14C in a special medium. An incubation period of six hours was sufficient for the radioactivity to reach an equilibrium between the fruit tissue and the incubation medium. Analyses of the fruit tissue and the medium for acidic and neutral metabolites of tryptophan indicated the existence of indolic products. The auxin indole-3-acetic acid (IAA) was identified among the products by dry column chromatography and biological assay. Other acidic metabolites included indolepyruvic acid and an unidentified material. Neutral metabolites included indolealdehyde, indoleacetaldehyde, and two unidentified compounds. Biological activity in the Avena curvature test was obtained from extracted compounds which corresponded to IAA and indolepyruvic acid in the acidic fraction and indoleacetaldehyde in the neutral fraction. Radioactive tryptophan was also found in both the acidic and the neutral fractions due to its amphoteric nature. The experiment demonstrated the conversion of tryptophan to its indolic metabolites, including indole-3-acetic acid, in this Citrus tissue. 相似文献
19.
A quantitative study of indole-3-acetic acid (IAA) turnover, and the contribution of tryptophan-dependent and tryptophan-independent
IAA-biosynthesis pathways, was carried out using protoplast preparations and shoot apices obtained from wild-type and transgenic,
IAA-overproducing tobacco (Nicotiana tabacum L.) plants, during a phase of growth when the level of endogenous IAA was stable. Based on the rate of disappearance of [13C6]IAA, the half-life of the IAA pool was calculated to be 1.1 h in wild-type protoplasts and 0.8 h in protoplasts from the
IAA-overproducing line, corresponding to metabolic rates of 59 and 160 pg IAA (μg Chl)−1 h−1, respectively. The rate of conversion of tryptophan to IAA was 15 pg IAA (μg Chl)−1 h−1 in wild-type protoplasts and 101 pg IAA (μg Chl)−1 h−1 in protoplasts from IAA-overproducing plants. In both instances, IAA was metabolised more rapidly than it was synthesised
from tryptophan. As the endogenous IAA pools were in a steady state, these findings indicate that IAA biosynthesis via the
tryptophan-independent pathway was 44 pg IAA (μg Chl)−1 h−1 and 59 pg IAA (μg Chl)−1 h−1, respectively, in the wild-type and transformed protoplast preparations. In a parallel study with apical shoot tissue, the
presumed site of IAA biosynthesis, the rate of tryptophan-dependent IAA biosynthesis exceeded the rate of metabolism of [13C6]IAA despite the steady state of the endogenous IAA pool. The most likely explanation for this anomaly is that, unlike the
protoplast system, injection of substrates into the apical tissues did not result in uniform distribution of label, and that
at least some of the [2H5]tryptophan was metabolised in compartments not normally active in IAA biosynthesis. This demonstrates the importance of using
experimental systems where labelling of the precursor pool can be strictly controlled.
Received: 18 January 2000 / Accepted 24 February 2000 相似文献
20.
Relationship of indole-3-acetic acid and tryptophan concentrations in normal and 5-methyltryptophan-resistant cell lines of wild carrots 总被引:1,自引:0,他引:1
Z. R. Sung 《Planta》1979,145(4):339-345
A 5-methyltryptophan(5-MT)-resistant cell line of wild carrot (Daucus carota L.), W001, that exhibited auxin-independent callus growth, was found to accumulate indole-3-acetic acid (IAA) and tryptophan (trp). Anthranilate-synthetase activity in W001 cell extract was less sensitive to feedback inhibition by trp than in the original 5-MT-sensitive cell lines. It is hypothesized that the resistant enzyme allowed more trp synthesis and accumulation which, in turn, affected the IAA concentration in the cell. Since carrot cultures cannot regenerate in the presence of exogenous auxin, the elevated IAA concentration in W001 may be responsible for its drastically reduced capacity to regenerate. The relationship between trp and IAA levels was further investigated by examining the effect of 2,4-dichlorophenoxy acetic acid (2,4-D) on the endogenous concentration of trp and IAA. In general, the IAA level was reduced but the trp concentration was elevated when 2,4-D was present in the culture medium.Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- IAA
indole-3-acetic acid
- 5-MT
5-methyltryptophan
- 5-MTr
5-MT-resistant
- 5-MTs
5-MT-sensitive
- trp
tryptophan 相似文献