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Metabolism of Exogenous Indoleacetic Acid to Its Amide Conjugates in Cucumis sativus L.

Incubation of hypocotyl segments of light-grown Cucumis sativus L. in 0.1 millimolar 3-indoleacetic acid for 16 hours led to the formation of indoleacetylaspartate and indoleacetylglutamate. There was no evidence for the formation of other conjugates of 3-indoleacetic acid with individual amino acids during the period from 4 to 48 hours of incubation. Indoleacetylglutamate reached its maximum concentration after about 4 hours of incubation and indoleacetylaspartate after about 8 hours. These levels remained unchanged for at least 40 hours. Indoleacetylaspartate caused small increases in cucumber hypocotyl segment growth at high concentrations, 1 millimolar being more effective than 0.1 millimolar.

Journal Article↗

Bound indoleacetic Acid in Avena coleoptiles.

When C(14) carboxyl indoleacetic acid (IAA) is transported through Avena coleoptile sections a fraction of the activity becomes bound. The nature of this bound IAA has been investigated. Upon extraction with solvents and chromatography a substance having the R(F) of IAA in 4 solvents was detected. No evidence could be found for the formation of indoleacetyl conjugates. In pea stem sections subjected to a similar experimental regime good evidence was obtained for the occurrence of conjugates. When IAA was supplied exogenously to coleoptile sections floating in solutions the occurrence of conjugates was shown to be dependent on the presence of the primary leaf. In its absence no conjugates could be detected.On grinding coleoptile sections and subsequent centrifugation at 240 x g the radioactivity was found to be in the tissue fraction as opposed to the supernatant. The radioactivity cannot be removed from the tissue by extraction with water, buffer solution or treatment with ribonuclease. It is readily removed by 10% urea, crystalline trypsin and chymotrypsin. It is therefore concluded that IAA becomes bound to a protein. Bound IAA does not appear to be able to cause growth in Avena coleoptile sections.

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Induction of indoleacetic Acid synthetases in tobacco pith explants.

Formation of indoleacetic acid synthetases in tobacco pith explants was determined by following the growth of tissue cultures under conditions of indole-3-acetic acid (IAA) deprivation and by measuring the enzymatic conversion of tryptophan to IAA in the cultures. The pith explants obtained from the parent plant (Nicotiana glauca) and from basal regions of the tumor-prone hybrid (N. glauca x N. langsdorffii) both show a requirement for exogenous IAA for growth initiation in culture. The parent pith requires the constant presence of added IAA for continued growth, but hybrid pith, after initial treatment with IAA, will grow without further additions. IAA synthetases are detected in the cell homogenates of hybrid pith explants cultured with either continuous or initial IAA addition. These observations indicate that IAA may induce its own production. In contrast, IAA synthetases are not found in the parent pith under comparable culture conditions. Besides IAA, nonhormonal compounds such as indole and tryptophan are also capable of stimulating growth of hybrid pith, possibly through the induction of IAA synthetases needed for IAA formation. Indole and tryptophan are, however, inactive in growth promotion of the parent pith. These results suggest that the genomic expression of IAA synthetase formation is more stringently controlled in N. glauca than in the tumorprone hybrid.

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Early effects of boron deficiency on indoleacetic Acid oxidase levels of squash root tips.

The indoleacetic acid (IAA) oxidase activity of root tips of boron-sufficient, -deficient, recovering, and IAA-treated boron-sufficient squash plants (Cucurbita pepo L.) was determined. Apical and subapical root sections displayed an increase in IAA oxidase activity between 6 and 9 hours after boron was withheld, and after 24 hours the activity of the apical sections showed a 20-fold increase over +B controls. Root elongation of -B plants was inhibited before an increase in oxidase activity could be detected. Roots of plants subjected to 12 hours of -B treatment and then transferred to +B treatment for recovery regained normal elongation rates and oxidase activity within 18 to 20 hours. IAA treatment of +B plants increased IAA oxidase activity of apical and subapical root sections and also inhibited root elongation and caused symptoms similar to -B treatments.These results have demonstrated the earliest enzymic change for intact boron-deficient plants. The results are in agreement with the theory that boron deficiency symptoms may be the result of supraoptimal endogenous levels of IAA. These high levels of IAA may inhibit cell division and lead to an induction of the IAA oxidase enzyme.

Journal Article↗

Responses of Avena coleoptiles to suboptimal fusicoccin: kinetics and comparisons with indoleacetic Acid.

Proton excretion induced by optimal concentrations of indoleacetic acid (IAA) and fusicoccin (FC) differs not only in maximum rate of acidification but also in the lag before onset of H(+) excretion and in sensitivity to cycloheximide. Because these differences might simply be a consequence of the difference in rate of proton excretion, FC and IAA have now been compared using oat coleoptiles (cv. Victory) under conditions where the rates of acidification are more similar, i.e. suboptimal FC versus optimal IAA. As the concentration of FC is reduced, the rate of H(+) excretion decreases, the final equilibrium pH increases, and the lag before detectable acidification increases up to 7-fold. This enhanced lag period is not primarily a consequence of wall buffering, inasmuch as it persists when a low concentration of FC is added to sections which were already excreting H(+) in response to IAA. An extended lag also occurs, upon reduction of FC levels, in the hyperpolarization of the membrane potential, before enhancement of O(2) uptake and before the increased rate of Rb(+) uptake. The presence or absence of a lag is not a distinguishing feature between FC and IAA actions on H(+) excretion and cannot be used to discriminate between their sites of action. In contrast, the insensitivity of FC-induced H(+) excretion to cycloheximide, as compared with the nearly complete inhibition of this auxin effect by cycloheximide, persists even at dilute concentrations of FC. This seems to be a basic difference in H(+) excretion by IAA and FC.

Journal Article↗

Fluorescence polarization immunoassay of urinary 5-hydroxy-3-indoleacetic acid.

Fluorescence polarization immunoassay of 5-hydroxy-3-indoleacetic acid in urine is described and compared with liquid chromatography (electrochemical detection) and colorimetry. Reports of in-house performance data and results of clinical trials are included to emphasize the usefulness of the assay for routine work.

Chromatography, High Pressure Liquid↗

Cerebrospinal fluid indoleacetic acid in autistic subjects.

Cerebrospinal fluid (CSF) levels of the tryptamine metabolite, indoleacetic acid (IAA), have been measured in groups of autistic and control subjects. No significant difference was seen in group mean (+/- SEM) levels of CSF IAA (autistics 5.53 +/- 0.47 ng/ml, N = 10). The finding indicates that central metabolism of the behaviorally active trace amine tryptamine is probably normal in autism.

Adolescent↗

Indoleacetic Acid and Abscisic Acid Antagonism: II. On the Phytochrome-Mediated Attachment of Barley Root Tips on Glass.

The effects of low concentrations (10 nm or less) of indoleacetic acid and abscisic acid on the phytochrome-mediated attachment of barley root (Hordeum vulgare L., var. Compana) tips on glass have been found to be opposite to those reported for mung bean root tips. Unlike their effects on mung bean root tips, indoleacetic acid causes attachment while abscisic acid causes detachment of barley root tips. Both hormones specifically and rapidly counteract the action of the other. No effect on the photoreversibility of attachment was observed when several gibberellins, cytokinins, and ethylene were added to the assay medium over a wide concentration range.

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Occurrence of 4-cl-indoleacetic Acid in broad beans and correlation of its levels with seed development.

Seeds and leaves of Vicia faba L. contain considerable amounts of 4-Cl-indoleacetic acid as determined by gas-liquid chromatography and gas-liquid chromatography-mass spectrometry. The unchlorinated auxin could not be detected in either leaves or seeds at different stages of development. The developmental stages of fruits and seeds were characterized by the rate of straight growth of pods, the rate of fresh weight increase, and the development of the endosperm. The peak of 4-Cl-indoleacetic acid content per seed coincides with the maximum rate of water accumulation in the seeds.

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Role of Pseudomonas putida indoleacetic acid in development of the host plant root system.

Many plant-associated bacteria synthesize the phytohormone indoleacetic acid (IAA). While IAA produced by phytopathogenic bacteria, mainly by the indoleacetamide pathway, has been implicated in the induction of plant tumors, it is not clear whether IAA synthesized by beneficial bacteria, usually via the indolepyruvic acid pathway, is involved in plant growth promotion. To determine whether bacterial IAA enhances root development in host plants, the ipdc gene that encodes indolepyruvate decarboxylase, a key enzyme in the indolepyruvic acid pathway, was isolated from the plant growth-promoting bacterium Pseudomonas putida GR12-2 and an IAA-deficient mutant constructed by insertional mutagenesis. The canola seedling primary roots from seeds treated with wild-type P. putida GR12-2 were on average 35 to 50% longer than the roots from seeds treated with the IAA-deficient mutant and the roots from uninoculated seeds. In addition, exposing mung bean cuttings to high levels of IAA by soaking them in a suspension of the wild-type strain stimulated the formation of many, very small, adventitious roots. Formation of fewer roots was stimulated by treatment with the IAA-deficient mutant. These results suggest that bacterial IAA plays a major role in the development of the host plant root system.

Amino Acid Sequence↗

Elevation of serum indoleacetic acid levels in rats with experimental liver failure.

Changes in the serum levels of indoleacetic acid (IAA) of liver-injured rats induced with an injection of carbontetrachloride (CCl4) or D-galactosamine (GalN) were investigated. IAA showed a marked increase of 61.8% and 102.6% in CCl4 and GalN treated rats, respectively. The elevation of IAA was observed only in the early stages of hepatic failure.

Alanine Transaminase↗

Colorimetric assay of urinary 5-hydroxy-3-indoleacetic acid.

We describe a method for 5-hydroxy-3-indoleacetic acid in urine based on its reaction with nitrosonaphthol in the presence of mercaptoethanolamine, an odorless compound. Mercaptoethanolamine shifts the color maximum from 540 nm to 640 nm, intensifies the color and discharges the color of interfering substances. The method is rapid and free from interferences.

Acetaminophen↗

Indoleacetic Acid and the synthesis of glucanases and pectic enzymes.

Indoleacetic acid (IAA) and/or inhibitors of DNA, RNA or protein synthesis were added to the apex of decapitated seedlings of Pisum sativum L. var. Alaska. At various times up to 4 days, enzymic protein was extracted from a segment of epicotyl immediately below the apex and assayed for its ability to hydrolyse polysaccharides or their derivatives. With the exception of amylase, the total amounts per segment of all of the tested enzymes increased due to IAA treatment. The development of beta-1,4-glucanase (cellulase) activity per unit of protein or fresh weight proceeded according to a typical sigmoid induction curve. Pectinase was formed for about 2 days in control segments and IAA treatment resulted in continued synthesis for at least another 2 days provided cell division took place. beta-1,3-glucanase and pectinesterase activities were only enhanced by IAA to the extent that total protein levels increased. Reaction mechanisms for these effects and functions for the enzymes during growth are discussed.

Journal Article↗

Effects of rutin on vegetative growth of mung bean (Vigna radiata) seedlings and its interaction with indoleacetic acid.

Rutin is one of the natural flavonoids in plants. Effects of rutin and indoleacetic acid (IAA) on vegetative growth of mung bean seedlings were studied and the contents of rutin and IAA in the hypocotyls of the seedlings were measured. Mung bean seedlings absorbed rutin through roots, grew more vigorously under light at lower rutin concentrations (20-60 microg/mL) with longer shoot axis (epicotyl+hypocotyl) and primary root and more lateral roots. The seedlings exhibited inhibition of elongation of the hypocotyls and radicles when grown in the dark at rutin concentrations of 20-100 microg/mL or under light at rutin concentrations of 80-100 microg/mL. Seedlings under light absorbed more rutin than those in the dark at rutin concentrations of 60-80 microg/mL and absorbed less rutin at concentrations lower than 40 microg/mL or near 100 microg/mL. IAA (>0.1 microg/mL) treatment promoted rutin accumulation and inhibited elongation of shoot axis and primary roots. The rutin content in hypocotyls reached the highest level when IAA (0.5 microg/mL) and rutin (40 microg/mL) were added to the medium. Rutin repressed endogenous IAA accumulation in hypocotyls of the etiolated seedlings and inhibited absorption of exogenous IAA from media to seedlings.

Fabaceae↗

Nucleotide sequences of the Pseudomonas savastanoi indoleacetic acid genes show homology with Agrobacterium tumefaciens T-DNA.

We report the nucleotide sequences of iaaM and iaaH, the genetic determinants for, respectively, tryptophan 2-monooxygenase and indoleacetamide hydrolase, the enzymes that catalyze the conversion of L-tryptophan to indoleacetic acid in the tumor-forming bacterium Pseudomonas syringae pv. savastanoi. The sequence analysis indicates that the iaaM locus contains an open reading frame encoding 557 amino acids that would comprise a protein with a molecular weight of 61,783; the iaaH locus contains an open reading frame of 455 amino acids that would comprise a protein with a molecular weight of 48,515. Significant amino acid sequence homology was found between the predicted sequence of the tryptophan monooxygenase of P. savastanoi and the deduced product of the T-DNA tms-1 gene of the octopine-type plasmid pTiA6NC from Agrobacterium tumefaciens. Strong homology was found in the 25 amino acid sequence in the putative FAD-binding region of tryptophan monooxygenase. Homology was also found in the amino acid sequences representing the central regions of the putative products of iaaH and tms-2 T-DNA. The results suggest a strong similarity in the pathways for indoleacetic acid synthesis encoded by genes in P. savastanoi and in A. tumefaciens T-DNA.

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