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Calcium ion dependency of ethylene production in segments of primary roots of Zea mays.

We investigated the effect of Ca2+ on ethylene production in 2-cm long apical segments from primary roots of corn (Zea mays L., B73 x Missouri 17) seedlings. The seedlings were raised under different conditions of Ca2+ availability. Low-Ca and high-Ca seedlings were raised by soaking the grains and watering the seedlings with distilled water or 10 mM CaCl2, respectively. Segments from high-Ca roots produced more than twice as much ethylene as segments from low-Ca roots. Indoleacetic acid (IAA; 1 micromole) enhanced ethylene production in segments from both low-Ca and high-Ca roots but auxin-induced promotion of ethylene production was consistently higher in segments from high-Ca roots. Addition of 1-aminocyclopropane-1-carboxylic acid (ACC) to root segments from low-Ca seedlings doubled total ethylene production and the rate of production remained fairly constant during a 24 h period of monitoring. In segments from high-Ca seedlings ACC also increased total ethylene production but most of the ethylene was produced within the first 6 h. The data suggest that Ca2+ enhances the conversion of ACC to ethylene. The terminal 2 mm of the root tip were found to be especially important to ethylene biosynthesis by apical segments and, experiments using 45Ca2+ as tracer indicated that the apical 2 mm of the root is the region of strongest Ca2+ accumulation. Other cations such as Mn2+, Mg2+, and K+ could largely substitute for Ca2+. The significance of these findings is discussed with respect to recent evidence for gravity-induced Ca2+ redistribution and its relationship to the establishment of asymmetric growth during gravitropic curvature.

Amino Acids↗

[Effects of Cd2+ on seedling growth and phytohormone contents of Glycine max].

A laboratory incubation experiment was conducted to study the effects of different Cd2+ concentrations on seedling growth and phytohormone contents of Glycine max through determining some physiological and biochemical indexes. The results showed as follows: (1) Different Cd2+ concentrations inhibited the synthesis of indoleacetic acid (IAA) and gibberellins (GA3) in roots and stimulated the synthesis of zeatin (Z) and abscisic acid (ABA) not only in roots but also in aerial parts of Glycine Max. Cd2+ stimulated the synthesis of IAA and GA3 in aerial parts at lower concentrations, but inhibited the synthesis of IAA and GA3 at higher concentrations. (2) Cd2+ stress for 84h increased the root vitality and the contents of chlorophyll a and b, but no significant difference was found in carotinoid contents compared with the control. (3) Cd2+ stress decreased POD activities and MAD contents atlower concentrations and increased POD activities and MAD contents at higher concentrations. With increasing Cd2+ concentrations, this increase became significant gradually. (4) Cd2+ stress with lower concentrations stimulated the physiological activity of Glycine max seedling in a short time. When Cd2+ concentration was 0.50 mg/L, the root vitality, IAA and GA3 contents of Glycine max were the highest, and POD activity was the lowest.

Abscisic Acid↗

The Role of Auxin and Gibberellin in Controlling Lignin Formation in Primary Phloem Fibers and in Xylem of Coleus blumei Stems.

The hypothesis that auxin (IAA) and gibberellic acid (GA(3)) control the formation of lignin is confirmed for the primary phloem fibers and for the secondary xylem in the stem of Coleus blumel Benth. Indoleacetic acid alone, or a combination of high IAA/low GA(3) (w/w), induced short phloem fibers with thick secondary walls, that contained lignin rich in syringyl units (high ratio of syringyl/guaiacyl). On the other hand, a combination of high GA(3)/low IAA (w/w), which promoted the differentiation of long phloem fibers with thin walls, decreased the relative content of the syringyl units (low syringyl/guaiacyl ratio). In the secondary xylem, these hormonal treatments yielded only slight changes in the noncondensed monomeric guaiacyl units, confirming the relative stability of the guaiacyl lignification pattern in this tissue. In the xylem, indoleacetic acid alone, or a combination of high IAA/low GA(3) induced lignin poor in syringyl units (low syringyl/guaiacyl ratio). A combination of high GA(3)/low IAA promoted a relatively slight increase in syringyl yield, indicating greater responsiveness of the syringyl lignification pattern to growth regulators. The possible functional and technological significance of our results is discussed.

Journal Article↗

Intermittent dystonia in Hartnup disease.

A 6-month-old girl developed intermittent dystonic posture of the legs and eczematous dermatitis without ataxia. Qualitative and quantitative urine amino acid testing confirmed the diagnosis of Hartnup disease. Cranial computed tomography, electroencephalogram, electromyogram/nerve conduction study, posterior tibial somatosensory evoked potentials, 24-hour electroencephalographic telemetry, and metrizamide myelogram were normal. Spinal fluid hydroxy-indoleacetic acid concentration was less than or equal to 2 S.D. of normal; oral tryptophan loading (70 mg/kg) resulted in a two-fold rise in cerebrospinal fluid 5-hydroxy-indoleacetic acid concentration. Tryptophan administered alone or with nicotinic acid failed to improve the dystonia; however, trihexyphenidyl (1-2 mg/kg/day) dramatically improved it. Hartnup disease should be considered in children with unexplained dystonia.

Brain Chemistry↗

Use of High Performance Liquid Chromatography for the Determination of Endogenous Hormone Levels in Solanum tuberosum L. Subjected to Carbon Dioxide Enrichment of the Root Zone.

Changes in cytokinin, auxin, and abscisic acid levels were determined by high performance liquid chromatography following CO(2) enrichment of the root zone. Both trans zeatin and zeatin riboside significantly decreased in the roots 6 days after treatment with CO(2). In the leaves zeatin riboside increased whereas trans zeatin showed no significant difference from the control. Indoleacetic acid in the leaves increased dramatically for 6 days following CO(2) treatment; however, indoleacetic acid levels in the roots increased slightly after 2 days and remained higher than the control. Abscisic acid increased shortly after CO(2) treatments; after 6 days this difference was no longer evident.

Journal Article↗

Effect of T-2 toxin on brain biogenic monoamines in rats and chickens.

Two experiments were conducted to determine the effect of T-2 toxin on brain biogenic monoamines and their metabolites. Male rats (180 g) and cockerels (28 day, 300 g) were orally dosed with T-2 toxin at 2.5 mg kg-1 body weight. In the first experiment, whole brains were collected 2, 6, 12, 24 and 48 h postdosing and analyzed for monoamines by high performance liquid chromatography with electro-chemical detection. T-2 toxin did not influence whole brain concentrations of monoamines in either species. In the second experiment, brains were collected 24 h postdosing, dissected into five brain regions, and analyzed for monoamines. T-2 toxin treatment resulted in increased serotonin and 5-hydroxy-3-indoleacetic acid in all brain regions of the rat. However, this was not seen in poultry where T-2 toxin treatment resulted in an increase in 5-hydroxy-3-indoleacetic acid, no alteration in serotonin concentration and a decrease in regional norepinephrine and dopamine concentrations. These results suggest that T-2 toxin influences brain biogenic amine metabolism and that there is an intraspecies difference in the central effects of this mycotoxin.

3,4-Dihydroxyphenylacetic Acid↗

Auxin-gibberellin interaction in apical dominance.

Indoleacetic acid and gibberellic acid were added to decapitated light-grown ;Alaska' pea seedlings as substitutes for the intact apex in the control of apical dominance. Of various concentrations and combinations tried, a combination of 1% indoleacetic acid + 1% gibberellic acid was the most inhibiting to side bud growth. The greatest degree of hormonally induced side bud inhibition was achieved when seedlings were deprived of soil nutrients.

Journal Article↗

Two components of auxin transport.

The transport of indoleacetic acid-1-(14)C out of sunflower stem sections has been analyzed by a compartmental analysis procedure in which the radioactivity moving out of the tissue (log per cent) is plotted against time. The analysis indicates that indoleacetic acid is transported via a fast transport system (t((1/2)) of about 30 minutes) and a slow transport system (t((1/2)) about 10 hours). While we do not know the sources of these two pools, by analogy with ion transport studies, the fast efflux is characteristic of transport from the cytoplasm across the plasmalemma and the slow efflux is characteristic of transport across the tonoplast and thus out of the vacuole. Both components of transport are inhibited by 2,3,5-triiodobenzoic acid.

Journal Article↗

Effect of glyphosate on auxin transport in corn and cotton tissues.

Basipetal auxin transport in 6-day-old dark-grown corn coleoptiles was severely inhibited by increasing levels of glyphosate applied during the transport period.The velocity of basipetal transport of [(14)C]indoleacetic acid in hypocotyls from 7-day-old cotton seedlings was significantly reduced when sublethal doses of glyphosate [N-(phosphonomethyl)glycine] were applied to the cotyledonary leaves 24 hours before transport measurement. Simultaneous application of glyphosate and indoleacetic acid during transport measurement had no effect on basipetal transport in cotton hypocotyl sections.Slowing of transport was inversely proportional to the dosage applied to both species.

Journal Article↗

Hormonal Regulation, and Intracellular Localization of a 33-kD Cationic Peroxidase in Excised Cucumber Cotyledons.

Ethylene enhanced chlorosis and levels of 33-kilodalton cationic peroxidase (33-CPO) in excised cucumber (Cucumis sativus L. cv ;Poinsett 76') cotyledons. Compared to other hormones, such as kinetin, indoleacetic acid, gibberellic acid, and abscisic acid, ethylene was the only effective promoter of 33-CPO synthesis. The hypothesis that peroxidase plays a role in chlorophyll degradation was tested by comparing levels of 33-CPO in cotyledons treated with compounds thought to either retard (kinetin, indoleacetic acid and gibberellic acid), or promote (abscisic acid and methyl jasmonate [MJ]) senescence. It was concluded that 33-CPO did not play a role in senescence since no direct correlation between chlorophyll content and 33-CPO was observed. MJ was as effective as ethylene in inducing senescence. However, ethylene did not appear to be involved in the action of MJ. Using immunocytochemistry, 33-CPO was found to be located primarily around starch grains and near the plasmalemma. High levels of 33-CPO were also found in cells destined to be vascular tissue.

Journal Article↗

Uptake and metabolism of serotonin by rat adrenal tissue in vitro.

After incubation of separate zones of rat adrenals with radioactively labelled serotonin--at concentrations at which the amine had been previously shown to stimulate aldosterone biosynthesis--the tissue radioactivity was several times higher in the capsular ("zona glomerulosa") than in the decapsulated portions of the glands. High doses of unlabelled serotonin diminished the accumulation of radioactivity, but only when added simultaneously with the tracer. In the course of the incubation, radioactive 5-hydroxy-3-indoleacetic acid, i.e. the deaminated metabolite of serotonin, rapidly appeared in the medium. Most of the radioactive material accumulated by the adrenal was recovered in the cytosol fraction and was no longer identical with serotonin, but consisted mainly of 5-hydroxy-3-indoleacetic acid and another, unidentified metabolite. Unaltered serotonin was found in capsular adrenal tissue extracts only when a monoamine oxidase inhibitor (nialamide) has been added to the incubation medium. Rapid uptake and metabolism thus appeared to be the predominant features of the interaction of incubated adrenal tissue with serotonin. These events may have obscured the possible binding of very small amounts of serotonin to receptor sites. The observed accumulation of radioactivity in adrenal tissue was mainly due to delayed outflow of intracellulary formed metabolite, since 5-hydroxy-3indoleacetic acid was neither taken up by incubated adreanls nor bound by adrenal cytosol. No evidence of serotonin storage in the zona glomerulosa was found in these studies.

Adrenal Glands↗

An excellent source of vegetative buds for use in plant hormone studies on apical dominance.

When studying the role of plant hormones in the control of growth at apical meristems, it is often difficult to obtain needed amounts of physiologically uniform buds. A source and method are described for obtaining sufficient quantities of large, uniform buds and for the treatment of the buds with indoleacetic acid and kinetin. Buds from the root system of Euphorbia esula L. were grown in Petri plates, with agar suspending the short root sections from which they emanate. Plant hormones are applied by their incorporation in the agar. The effect of various concentrations of indoleacetic acid and kinetin on bud growth was examined.

Journal Article↗

Identification and kinetics of accumulation of proteins induced by ethylene in bean abscission zones.

A two-dimensional gel electrophoresis system that combines a cationic polyacrylamide gel electrophoresis at pH near neutrality with sodium dodecyl sulfate-polyacrylamide gel electrophoresis was used to analyze the spectrum of basic polypeptides that accumulate in bean (Phaseolus vulgaris) abscission zones after treatment with ethylene. Results showed that, as abscission progressed, at least seven basic proteins accumulated in the abscission zone prior to the accumulation of 9.5 cellulase. Six of the seven proteins correspond to pathogenesis-related (PR) proteins. Among them, two isoforms of beta-1,3-glucanase and multiple isoforms of chitinase were identified. A 22 kilodalton polypeptide that accumulated to high levels was identified as a thaumatin-like protein by analysis of its N-terminal sequence (up to 20 amino acids) and its serological relationship with heterologous thaumatin antibodies. A 15 kilodalton polypeptide serologically related to PR P1 (p14) from tomato was identified as bean PR P1 (p14)-like protein. The kinetics of accumulation of glucanases, chitinases, thaumatin-like and PR P1 (p14)-like proteins during ethylene treatment were similar and they showed that PR proteins accumulated in abscission zones prior to the increase in 9.5 cellulase. Addition of indoleacetic acid, a potent inhibitor of abscission, reduced the accumulation of these proteins to a similar extent (60%). The synchronized accumulation of this set of PR proteins, early in the abscission process, may play a role in induced resistance to possible fungal attack after a plant part is shed. The seventh protein does not correspond to any previously characterized PR protein. This new 45 kilodalton polypeptide accumulated in abscission zones on exposure to ethylene concomitantly with the increase in 9.5 cellulase. Its N-terminal sequence (up to 15 amino acids) showed some homology with the amino terminal sequence of chitinase. Polyclonal antibodies against chitinase recognized the 45 kilodalton polypeptide, but polyclonal antibodies against the 45 kilodalton protein recognized chitinase weakly. When abscission was inhibited by addition of indoleacetic acid, the accumulation of the 45 kilodalton protein was strongly inhibited (80%). This result suggests that the 45 kilodalton polypeptide may play a more direct role in abscission.

Journal Article↗

The role of ethylene in the senescence of oat leaves.

The evolution of ethylene, both from the endogenous source and from added 1-aminocyclopropane-1-carboxylic acid (ACC), has been followed in close relationship with the senescent loss of chlorophyll from seedling oat leaves. In white light, where chlorophyll loss is slow, the ethylene evolution increases slowly at first, but when the loss of chlorophyll becomes more rapid, ethylene evolution accelerates. CoCl(2) inhibits this increase and correspondingly maintains the chlorophyll content, with an optimum concentration of 10 micromolar. The rapid rate of chlorophyll loss in the dark is slightly decreased by 3-aminoethoxyvinyl glycine (AVG), by cobalt, and slightly stimulated by ACC. The slower chlorophyll loss in white light, however, is almost completely inhibited by silver ions, greatly decreased by cobalt and by AVG, and strongly increased by ACC. Since the chlorophyll loss is accompanied by proteolysis, it represents true senescence. Chlorophyll loss in light is also strongly antagonized by CO(2), 1% CO(2) giving almost 50% chlorophyll maintenance in controls, while in the presence of added ACC or ethylene gas, the chlorophyll loss is 50% reversed by about 3% CO(2). The ethylene system in leaves is thus more sensitive to CO(2) than that in fruits. Indoleacetic acid also clearly decreases the effect of ACC. It is shown that kinetin, CO(2), Ag(+), and indoleacetic acid, all of which oppose the effect of ethylene, nevertheless increase the evolution of ethylene by the leaves, and it is suggested that ethylene evolution may, in many instances, mean that its hormonal metabolism is being prevented.Abscisic acid somewhat increases ethylene evolution also, but its action in promoting senescence in light is antagonized only partially by Ag(+), Co(2+), or AVG. For this and a number of other reasons it is concluded that ethylene and abscisic acid both independently control leaf senescence in the light.

Journal Article↗

Test for decreased serotonin/tryptophan metabolite ratios in abstinent alcoholics.

A segment of the population has a defect in the metabolism of tryptophan that causes a lowered concentration of serotonin in the central nervous system and indicates a predisposition towards chronic alcoholism. The metabolic defect in tryptophan metabolism is in the pathway between tryptophan and 5-hydroxyindoleacetic acid (5-HIAA) but not in the other pathways of tryptophan metabolism. A test using HPLC with amperometric detection was developed to detect the presence of an impaired serotonin metabolic pathway and therefore indicate a predisposition towards chronic alcoholism. The test used a ratio between 5-HIAA and two metabolites (indoleacetic acid and anthranilic acid) from the other pathways of tryptophan metabolism to indicate the presence or absence of an impaired serotonin metabolic pathway while correcting for variations in the concentration of urine.

Adult↗

Mechanism for transcriptional action of cyclic AMP in Escherichia coli: entry into DNA to disrupt DNA secondary structure.

Binding analysis with purified bacterial receptor distinguishes two structural domains in cyclic AMP (cAMP). The first, the cyclic phosphate and furanose, constitutes a binding domain. This region is bound tightly to the receptor. The rest of cAMP is not bound; the adenine moiety of cAMP is exposed. Unlike binding, activity of cAMP requires the adenine moiety. To be active, cAMP must have in domain II the base adenine--specifically, its Watson--Crick atoms N-1 and N-6. Analysis of indoleacetic acid, a compound able to replace cAMP at the L-arabinose operon, indicates a similar distinction between binding and active domains. To be active, the indole must have substitution (carboxyl or amide) electronically comparable to the cAMP N-1 and N-6. On this basis, we propose a detailed mechanism for action of cAMP (or indoleacetic acid) in Escherichia coli. We propose that the exposed adenine of cAMP enters into the DNA. The adenine's N-1 and N-6 form hydrogen bonds to a thymine in DNA. This interaction destabilizes the DNA. It enhances transcription. Marked similarities indicate an identical mechanism for the steroid hormones in eukaryotes.

Cyclic AMP↗

Azido auxins: synthesis and biological activity of fluorescent photoaffinity labeling agents.

Three auxin analogs, 4-, 5-, and 6-azido-3-indoleacetic acid (4-N(3)-IAA, 5-N(3)-IAA, and 6-N(3)-IAA) have been synthesized for use as fluorescent photoaffinity labeling agents. The pK(a) values of these compounds (4-N(3)-IAA, 4.67; 5-N(3)-IAA, 4.65; 6-N(3)-IAA, 4.66; all +/- 0.04) are experimentally indistinguishable from the pK(a) of 3-indoleacetic acid (IAA, 4.69 +/- 0.04). The auxin activity of these IAA derivatives has been determined in several systems. In soybean, pea, and corn straight growth assays, all three analogs induce growth comparable to that caused by IAA. In the tobacco pith assay, all three analogs elicit a maximum increase in fresh weight at least 40 to 50% of that caused by IAA. Optimal growth is attained in the tobacco pith assay at slightly higher concentrations of 4-N(3)-IAA and 6-N(3)-IAA (30 micromolar) than required for IAA (10 micromolar); however, maximal growth is achieved at a slightly lower concentration of 5-N(3)-IAA (3 micromolar). The N(3)-IAAs, like IAA, are transported basipetally through tobacco pith tissue.

Journal Article↗

Lateral movement of auxin in phototropism.

Lateral movement of indoleacetic acid-1-(14)C in corn coleoptiles was measured as radioactivity moving laterally following unilateral application of the auxin. The data suggest that there is an endogenous lateral movement of auxin, and that phototropic stimulation of the coleoptile depresses lateral movement towards the light and enhances lateral movement away from the light. The lateral movement was found to be principally as indoleacetic acid. In experiments using sunflower hypocotyl sections, evidence is also presented to support the suggestion that lateral redistribution of auxin may be effected by a deflection of auxin around a barrier to basipetal transport.

Journal Article↗