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Relationship between Promotion of Xyloglucan Metabolism and Induction of Elongation by Indoleacetic Acid.

Auxin promotes the liberation of a xlyoglucan polymer from the cell walls of elongating pea (Pisum sativum) stem segments. The released polymer can be isolated from the polysaccharide fraction of the water-soluble portion of tissue homogenates, thus providing as assay for this kind of metabolism. Promotion of xyloglucan metabolism by auxin begins within 15 minutes of hormone presentation. The effect increases with auxin concentration in a manner similar to the hormone effect on elongation. However, the xyloglucan effect of auxin occurs perfectly normally when elongation is completely blocked by mannitol. Metabolic inhibitors and Ca(2+), on the other hand, inhibit auxin promotion of elongation and of xyloglucan metabolism in parallel. The results suggest that the changes in xyloglucan reflect the means by which auxin modifies the cell wall to cause elongation.

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Interactions of microtubule disorganizers, plant hormones, and red light in wheat coleoptile segment growth.

Growth response of coleoptile segments excised from 3-day-old seedlings of wheat (Triticum vulgare cv. Baart) to gibberellic acid, indoleacetic acid, and 2,4-dichlorophenoxyacetic acid, to red light, and to several microtubule disorganizers depends on the initial position of the excised segment in the intact coleoptile. Red light, 660 nm, stimulates the growth of the apical cells, but inhibits markedly the growth of the cells in the basal region of the coleoptile. The effects of red light are independent of sucrose, gibberellic acid, indoleacetic acid, and 2,4-dichlorophenoxyacetic acid, even though these substances themselves markedly affect the growth of the coleoptile segments. Concentractions of the microtubule disorganizers, vinblastine sulfate, cupric chloride, urea, and colchicine, which do not alter significantly the growth of the dark control apical segments, substantially repress the promotive effects of red light or auxin on the increase in length of the apical cells of the coleoptile. This suggests that stimulation by red light and by auxin involves microtubule production. Microtubule disorganizers repress the growth of elongating cells of the coleoptile, yet on the other hand, auxin and irradiation do not alter significantly the response of basal cells to the microtubule disorganizing agents. We hypothesized that light and growth regulators induce polymerization of nonaggregated microtubule subunits, resulting in faster growth.

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Translocation and Metabolism of Endosperm-Applied [2-C] Indoleacetic Acid in Etiolated Avena sativa L. Seedlings.

The role of free indole-3-acetic acid (IAA) in the endosperm of Avena sativa L. seedlings was investigated to determine its contribution to free IAA in the shoot. [2-(14)C]IAA was injected into the endosperm of darkgrown seedlings and the transport and metabolism of the [(14)C]-labeled compounds determined. It was concluded that translocation of free IAA directly from the endosperm is probably not a significant source of free IAA in the shoot, mainly because even small amounts of [(14)C]IAA introduced into the endosperm were rapidly metabolized. This suggested that, in Avena, free IAA does not normally exist in the liquid endosperm.

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Role of calcium in the polar secretion of indoleacetic Acid.

The rate of auxin transport in sunflower hypocotyls (Helianthus annuus L. cv ;Russian mammoth') or corn coleoptiles (Zea mays L. cv ;WF9 x 38') was less in seedlings grown in Ca-deficient medium than in controls. The rate of IAA transport depended on the concentration of Ca in the root medium up to 1 millimolar. Further increases in auxin transport were observed when the isolated segments were incubated in medium containing up to 30 millimolar Ca. We suggest that the rate of auxin transport in plant tissue is dependent on the pool of ionic Ca in the extracellular space.Segments from Ca-deficient seedlings exhibited a high specific requirement for Ca(2+) in auxin transport. Magnesium, strontium, and several other divalent cations tested for their ability to replace Ca(2+) in restoring auxin transport showed no effect; partial replacement by lanthanum was observed.Auxin transport, or auxin flux through the segment, which is the result of IAA secretion by individual cells, was reduced in the low Ca(2+) segments due both to lowered velocity and to reduced capacity of transport. The requirement for Ca(2+) in the secretion of auxin is believed to be equivalent to the phenomenon observed in animal cell secretion, where the influx of Ca(2+) serves as a link between an external stimulus and the secretion response.

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Promotion of Stomatal Opening by Indoleacetic Acid and Ethrel in Epidermal Strips of Vicia faba L.

Indole-3-acetic acid (IAA), at concentrations of 0.01 to 1.0 millimolar, and ethephon (0.3% v/v Ethrel) promote stomatal opening when applied to epidermal peels of Vicia faba L. in light or dark. The effect of ethylene is seen by 30 minutes and maximal opening (over two times that of untreated controls) occurs after only 60 to 90 minutes in the light. Stomatal opening by IAA and Ethrel in both light and dark is prevented by 0.14 millimolar AgCl. It is suggested that the effect of added IAA, but not that of light, is linked to ethylene production. The possible role of ethylene in stomatal opening during fungal infection is discussed. The stomates of Vicia faba provide a new system to study the effects of ethylene on certain membrane-regulated processes.

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Quantitation of indoleacetic Acid conjugates in bean seeds by direct tissue hydrolysis.

Gas chromatography-selected ion monitoring-mass spectral analysis using [(13)C(6)]indole-3-acetic acid (IAA) as an internal standard provides an effective means for quantitation of IAA liberated during direct strong basic hydrolysis of bean (Phaseolus vulgaris L.) seed powder, provided that extra precautions are undertaken to exclude oxygen from the reaction vial. Direct seed powder hydrolysis revealed that the major portion of amide IAA conjugates in bean seeds are not extractable by aqueous acetone, the solvent used commonly for IAA conjugate extraction from seeds and other plant tissues. Strong basic hydrolysis of plant tissue can be used to provide new information on IAA content.

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Cytokinins and Differentiation Processes in Mercurialis annua: Genetic Regulation, Relations with Auxins, Indoleacetic Acid Oxidases, and Sexual Expression Patterns.

Cytokinins in apices of eight isogenic lines of Mercurialis annua were compared (high performance liquid chromatography-gas chromatography mass spectroscopy-computer system). These apices develop normal staminate or pistillate differentiation processes (sex series lines) or empty (sterile), semiempty (semisterile), and full anthers (restored fertile male) in the sterility series in which a pistillate line was constructed. Both series developed two different cytokinin pathways: trans-cytokinins characterized the sex series, whereas the cis pathway characterized the sterility series. Drastic changes in the trans pathway (0/250 nanograms trans-zeatin and 166/0 nanograms zeatin nucleotide) induced staminate/pistillate differentiations. Less drastic quantitative changes in the cis pathway induced sterility or restored fertility compared to normal fertile anthers (192 or 669 nanograms/traces). The action of the complete cis-pathway was morphologically effective in the sterility series when the ratio of cis to trans pathways was 1:2 or 1:1 instead of 1:3. A final diagram shows the action of each sex or sterility allele on the enzymes controlling specific metabolites in both pathways. The discussion provides insights on the regulation of cytokinin-auxin balances specific for each kind of reproductive differentiation.

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Western blot analysis of cereal grain prolamins using an antibody to carboxyl-linked indoleacetic Acid.

A monoclonal antibody raised against carboxyl-linked IAA was used in Western blot analysis of storage proteins from kernels of Avena sativa, Pennisetum americanum, Sorghum bicolor, and Zea mays. IAA or an IAA-like molecule is associated with the ethanolsoluble protein fraction of the seed. Western blotting of commercial zein, the major storage protein of maize, along with physicochemical evidence reported by Leverone et al. ([1991] Plant Physiol, 96: 1070-1075) indicated that IAA is linked with this prolamin. Results suggest that an IAA-prolamin association may be widespread throughout the Poaceae.

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Oxidized oligogalacturonides activate the oxidation of indoleacetic Acid by peroxidase.

Partial hydrolysis of polygalacturonic acid with a purified alpha-1,4-endopolygalacturonase yielded oligogalacturonides and trace amounts of a series of modified oligogalacturonides. Three of the minor products were isolated and identified as oxidized oligogalacturonides possessing termini of galactaric acid. Oxidation of indole-3-acetic acid by peroxidases was activated by oxidized oligogalacturonides but not by normal analogs.

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