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PubMed1967

Modification of apparent phytochrome synthesis in pisum by inhibitors and growth regulators.

The repeated exposure of Pisum (pea) plants to red light brings into operation an apparent synthesis of phytochrome which is not observed in material kept in the dark. This process shows some temperature compensation but has an optimum at 26 degrees ; it is irreversibly inhibited by 10(-4)m cycloheximide and 10 mug/ml actinomycin D. It is also inhibited by the auxins indoleacetic acid, naphthalene acetic acid and 2,4-dichlorophenoxyacetic acid at 10(-4)m but in these cases the inhibition is completely reversed when the auxin is washed out of the tissue. Antiauxins 2,4,6-trichlorophenoxyacetic acid and p-chlorophenoxy isobutyric acid, while strongly inhibiting growth have little effect on apparent synthesis. Other growth regulators and the precursor of tetrapyrrole synthesis, delta-aminolevulinic acid, have no consistent effect on the process, but 3 x 10(-4)m cobalt (II) nitrate is inhibitory. The capacity for apparent synthesis decreases as the cells approach maturity. The results may be explained by either de novo synthesis of phytochrome, or by a transformation process resembling in some respects the dark reversion of Pfr to Pr. The physiological role of apparent synthesis is suggested.

Journal Article↗
PubMed1967

Inhibition of polar auxin transport by ethylene.

Applied ethylene influences the growth of etiolated pea stem sections cut from untreated plants, but has no effect on (14)C-indoleacetic acid uptake, polar transport or destruction. However, the capacity of the polar auxin transport system is markedly reduced in sections cut from plants grown in ethylene, while the velocity of auxin transport is unchanged under these conditions. Inhibition of the polar transport system by ethylene could underlie certain responses in which the gas produces symptoms of auxin deficiency.

Biological Transport↗
PubMed1968

Studies on Auxin Protectors. IV. The Effect of Manganese on Auxin Protector-I of the Japanese Morning Glory.

Auxin protector-I of the Japanese morning glory is inactivated by manganese. Experiments carried out in vitro indicate that in the absence of oxygen the manganic, but not the manganous, ion rapidly inactivates the protector. It is clear from these, and other data described in this report, and the results of other workers, that in the presence of oxygen, manganese accelerates auxin inactivation by means of 2 separate and distinct mechanisms: 1) manganese catalyzes the oxidation of auxin protectors, and 2) following the inactivation of the protectors, or in the absence of protectors, accelerates the oxidation of indoleacetic acid by endogenous peroxidases.

Journal Article↗
PubMed1968

The auxin induced curvature of citrus petals.

The curvature of citrus petals was studied, using petals implanted in an agar-sucrose medium. Applications of indoleacetic acid and p-chlorophenoxyisobutyric acid and determinations of endogenous auxins indicate that the bending of citrus petals is an auxin-mediated process.

Journal Article↗
PubMed1968

Influence of Substrate and Tissue Manganese on the IAA-Oxidase System in Cotton.

Tissue manganese was found to influence the indoleacetic acid (IAA) system of cotton over a wide range of concentrations. The cofactor and inhibitor activities of the IAA-oxidase system were affected as the concentration of manganese in the tissue was varied. Maximum inhibitor activity was found in leaf extracts from the plants grown in 0.5 mg/l manganese (Hoagland's level). The inhibitor activity decreased in the leaf extracts of plants grown at concentrations of manganese either higher or lower than 0.5 mg/l. Abnormally high IAA-oxidase activity was found in the leaves of plants grown in deficient levels of manganese (<0.0005, 0.005 mg/l) and the extracts from plants in the <0.0005 mg/l Mn treatment showed IAA-oxidase cofactor activity.

Journal Article↗
PubMed1968

Relationships between the development of adventitious roots and the biosynthesis of anthocyanins in first internodes of sorghum.

The initiation and subsequent growth of adventitious roots in excised first internodes of Sorghum vulgare var. Wheatland milo were studied to determine the effect of these processes on anthocyanin biosyntheses. Segmentation of the internodes inhibited both adventitious root growth and accumulation of cyanidin equally in all segments; these results can be interpreted as a common requirement for bidirectional longitudinal transport. The presence of the coleoptile, especially in the absence of the base of the internode, inhibited the growth of the roots, but increased the number of root initials. High intensities of white and blue light which induced cyanidin synthesis slightly decreased adventitious root growth. Anaerobic conditions produced by solution infiltration strongly inhibited the growth of adventitious roots and greatly increased the accumulation of apigeninidin and luteolinidin. Addition of indoleacetic acid, kinetin and cofactors such as pyridoxine produced effects on the initiation and subsequent growth of these roots similar to those effects reported in the literature. But unlike root formation in hypocotyls, the initiation of adventitious roots in Sorghum internodes was not always directly correlated with the accumulation of anthocyanins, and the subsequent growth of these roots was frequently inversely correlated with some of the anthocyanin biosyntheses. The possible nature of these correlations is discussed. Comparisons are made with related Sorghum lines and mutants.

Journal Article↗
PubMed1969

IAA Oxidase Inhibitors from Normal and Mutant Maize Plants.

Extracts of maize (Zea mays L.) plants contain substances which, in vitro, inhibit an indoleacetic acid (IAA) oxidase enzyme from maize. The extracts can be freed of inhibitors by dialysis or by passage through columns of polyvinylpyrrolidone powder. Inhibitor-free extracts contain an IAA oxidase enzyme which requires a phenolic co-factor and is stimulated by Mn(2+).IAA oxidase inhibitor and total phenol levels were compared for normal maize and for the maize mutant Knotted (Kn). In plants up to 18 days old the level of heat stable, water soluble IAA oxidase inhibitors increases with increasing dosage of the Kn allele. Increased inhibitor content is accompanied, but not paralleled, by increased content of total phenols. Although several inhibitors are present in crude extracts, most inhibition can be attributed to 1 compound. This compound is not destroyed by horseradish peroxidase in the absence of IAA. At pH 3.5 it is not extracted into ether, but it is rendered ether-extractable by incubation in 2 n KOH for 5 hr at room temperature. This compound is tentatively identified as an ester of ferulic acid and some unknown moiety.

Journal Article↗
PubMed1969

Abscission: role of cellulase.

Cellulase (beta-1,4-glucan-glucanohydrolase EC 3.2.1.4) activity increased during abscission and was localized in the cell separation layer of Phaseolus vulgaris L. cv. Red Kidney (bean), Gossypium hirsutum L. cv. Acala 4-42 (Cotton) and Coleus blumei Benth. Princeton strain (Coleus) abscission zone explants. Cellulase activity was optimum at pH 7, was reduced by one-half after heating to 55 degrees for 10 min, and was associated with the soluble components of the cell. Explants treated with aging retardants (indoleacetic acid, (6)N-benzyladenine, and coumarin), CO(2), actinomycin D or cycloheximide had less cellulase activity than untreated controls. Ethylene increased cellulase activity of aged explants after a 3-hr lag period but had no effect on cellulase activity of freshly excised explants. It was concluded that 1 of the roles of ethylene in abscission is to regulate the production of cellulase which in turn is required for cell separation.

Journal Article↗
PubMed1969

Phototropism and photoinhibition of basipolar transport of auxin in oat coleoptiles.

We have proposed that the lateral inequality of auxin associated with the phototropic response is a consequence of a light-induced impairment of basipolar transport of the hormone. If this is so, the dose-response curve for photoinhibition of auxin transport should resemble that for phototropism. Further, the wavelength dependencies of the transport and tropic response should also be similar. Oat coleopilles were irradiated equilaterally with white light and with broad- and narrow-spectral bands of blue light. Indoleacetic acid 2-(14)C was then applied apically to the intact coleoptile. Irradiation by the 3 sources inhibited basipolar transport of the auxin. The photoinhibition of transport increases with exposure, reaches a maximum at radiant energies depending on the source used, and then decreases. The dose response for transport inhibition matches that for phototropism. A correspondence in the spectral response of the 2 phenomena is also found for coleoptiles exposed to first positive energies at various wavelengths from 340 to 730 nm. We interpret these correlations as support for the hypothesis that phototropism is mediated by a photoinhibition of basipolar transport of auxin.

Journal Article↗
PubMed1970

Regulation of root growth by auxin-ethylene interaction.

A large portion of indoleacetic acid (IAA)-induced inhibition of excised root tips and virtually all such inhibition of intact roots are the result of IAA-dependent ethylene production. Under certain conditions an additional effect of IAA accounts for a small portion of the inhibition of excised root tips. Ethylene production in response to applied IAA is governed by the level of applied auxin found inside the root. Evidence is presented to confirm the participation of ethylene in the geotropic response of roots.

Journal Article↗
PubMed1970

Time course of auxin stimulations of growth.

Measurements of the time course of growth responses of corn coleoptile sections to pulses of auxin (10(-5)m indoleacetic acid) establish that the growth rate changes in a regular pattern around the auxin pulse: a latent phase of 12 to 15 minutes is followed by an acceleration of growth rate lasting 15 to 20 minutes, after which a fairly steady rate is maintained. When the auxin source is withdrawn, there is an after-effect of about 15 minutes followed by a decay of growth rate, which reaches 50% decay after a further 15 to 40 minutes. The decay phase appears to be a function of the transport of auxin out of the sections. The 50% decay of growth for single cells is estimated at 30 minutes from the time of withdrawal of an exogenous supply of auxin. The regulation of growth by auxin is rapidly imposed or dissipated as auxin enters and exits, respectively, suggesting a facile association and disassociation of auxin with a growth-limiting site in the cell. It is proposed that the growth-stimulated state is dissipated at once when the transportable auxin has passed out of the cell.

Journal Article↗
PubMed1970

Choice of rotation rate for the horizontal clinostat.

A series of nine rates of rotation of the clinostat were tested to determine optimal and acceptable conditions for simulating weightlessness in plants. Young seedlings of wheat (Triticum aestivum L.) developed roots and coleoptiles of equal lengths and with the same orientation angles over a range of rotation rates from 0.25 to 480 minutes per revolution. Rates from 0.25 to 3 minutes per revolution provided for maximal epinastic curvatures of leaves and branches of Coleus blumei Benth. except for a reduced mean curvature of branches at 0.25 minute per revolution, due probably to physical disturbances in their growth. Smaller epinastic curvatures developed in both leaves and branches rotated at 15 minutes per revolution or more slowly. Indoleacetic acid-2-(14)C was used for measurements of extractable radioactivity in determining the reason for smaller curvatures of Coleus branches and tomato (Lycopersicon esculentum Mill.) leaves at the rotation rate of 60 minutes per revolution than at 1 minute per revolution. The cause was determined to be movement of some auxin from the upper into the lower side of a plagiotropic leaf or branch under the transient influence of gravity during rotation on the slower clinostat. A rotation period of 1 to 3 minutes was found to be acceptable for most plants.

Journal Article↗
PubMed1970

Studies on Auxin Protectors: IX. Inactivation of Certain Protectors by Polyphenol Oxidase.

Protector-II (Pr-II) of the Japanese morning glory (Pharbitis nil Choisy) was inactivated by exposure to polyphenol oxidase. An unidentified protector in the same molecular weight range obtained from sunflower was also inactivated by this enzyme. Earlier speculations that protectors might be lipoprotein in nature were negated by the fact that neither lipase nor protease inactivated the protectors. The protectors were also not inactivated by incubating with alpha-amylase, DNase, or RNase. Catechol mimics Pr and is inactivated by polyphenol oxidase. The oxidation of catechol to o-quinone is accompanied by a loss of chromophores that absorb ultraviolet light and the appearance of a reddish brown color. Similarly, when the relatively low molecular weight auxin protectors (Pr-II class) were incubated with polyphenol oxidase, their oxidation was also frequently associated with the formation of brown color, and oxidation with H(2)O(2) caused a loss of ultraviolet-absorbing chromophores. The data indicate that auxin protectors contain o-dihydroxyphenolic groups at their active site.That o-dihydroxyphenols inhibit indoleacetic acid oxidation has been demonstrated by numerous workers. It is suggested that the high molecular weight auxin protectors and the phenolic compounds described by other authors comprise part of a metabolic system concerned with the regulation of peroxidase-catalyzed redox reactions.

Journal Article↗
PubMed1971

The regulation of cambial division and secondary xylem differentiation in xanthium by auxins and gibberellin.

Cambial division continued in decapitated Xanthium plants without concomitant xylem fiber differentiation. The application of indoleacetic acid to these plants did not affect the production of cambial derivatives or induce xylem fiber differentiation. When naphthaleneacetic acid was applied either to the second internode or to the stump of a lateral shoot, xylem fiber differentiation was induced in the newly formed cambial derivatives on the xylem side of the cambium in the stem. When naphthaleneacetic acid was applied unilaterally, xylem fiber differentiation was restricted to that side of the stem in the first internode and hypocotyl. Naphthaleneacetic acid also enhanced the production of cambial derivatives. Gibberellic acid enhanced cambial derivative production but did not affect the differentiation of xylem fibers. Similar numbers of cambial derivatives were produced in some naphthaleneacetic acid-treated plants in which xylem fiber differentiation was induced and in gibberellic acid-treated plants which did not differentiate xylem. When naphthaleneacetic acid was applied 72 hours after decapitation, the oldest of the cambial derivatives on the xylem side failed to develop into fibers although younger cells did. These results suggest that auxin has its direct effect on the induction of xylem differentiation rather than the induction of divisions prerequisite to differentiation.

Journal Article↗
PubMed1971

The Conversion of d-Tryptophan to l-Tryptophan in Cell Cultures of Tobacco.

d-Tryptophan was converted to l-tryptophan in tissue cultures of tobacco, in whole cells treated with dimethylsulfoxide, and in cell-free extracts treated by Sephadex G-25 filtration. Evidence was obtained that tryptophanase, tryptophan pyrrolase, and transaminase activities were not involved. The data were best explained by the presence of a tryptophan racemase as the enzyme catalyzing the reaction. The possible role of d-tryptophan in the biosynthesis of indoleacetic acid is discussed.

Journal Article↗
PubMed1971

Regulation by auxin of carbohydrate metabolism involved in cell wall synthesis by pea stem tissue.

Promotion of cell wall synthesis (from glucose) in pea (Pisum sativum) stem segments by indoleacetic acid (IAA) develops over a period of 1 to 2 hours and is comprised of a promotion of glucose uptake plus a promotion of the utilization of absorbed glucose. The effect of IAA resembles, in these and other respects, its effect on cell wall synthesis in oat coleoptile segments, but the pea system differs in not being inhibited by galactose or mannose, in involving considerably more isotope dilution by endogenous substrates, and in certain other respects.EFFECTOR INFLUENCES UPON AND TOTAL ACTIVITIES OF THE FOLLOWING ENZYMES OBTAINED FROM ETIOLATED PEA STEM SEGMENTS PRETREATED WITH OR WITHOUT IAA WERE EXAMINED: phosphoglucomutase, uridine diphosphate glucose (UDP-glucose) pyrophosphorylase, nucleoside diphosphokinase, UDP-glucose dehydrogenase, inorganic pyrophosphatase, hexokinase (particulate and soluble), and UDP-glucose-beta-1,4-glucan-glucosyl transferase (beta-glucan synthetase). The first three enzymes mentioned exhibit high activity relative to the flux in vivo, do not appear to show physiologically significant effector responses, and are concluded not to be control points. UDP-glucose dehydrogenase activity is regulated by UDP-xylose. Hexokinase is a potential control point but does not exhibit regulatory effects related to the IAA response. beta-Glucan synthetase is the only one of these enzymes with activity which is increased by treatment of tissue with IAA, and this may be responsible for the effect of IAA on wall synthesis.Assays of metabolite pools support the conclusion that stimulation of polysaccharide synthesis by IAA is due partly to changes in hexokinase reaction rate resulting from an increase in metabolic glucose pool size caused by increased glucose uptake, and partly to increased activity at the polysaccharide synthetase level.

Journal Article↗
PubMed1971

Effect of the Hypocotyl Hook on Chlorophyll Accumulation in Excised Cotyledons of Cucumis sativus L.

Hypocotyl hooks have been shown to influence greening in excised cucumber (Cucumis sativus) cotyledons. The properties of the lag phase are greatly affected by the presence or absence of the hook tissue. A 45-second light pretreatment followed by 4 hours of darkness is sufficient to remove the lag phase from cotyledons with hooks, while hookless cotyledons require 2 hours of continuous illumination followed by 1 hour of dark incubation to break the lag phase. The effect of hooks on cotyledon greening is enhanced if the hooks are shielded from light. Cutting off the hooks after lag phase removal caused a marked decrease in chlorophyll accumulation in the cotyledons. These observations may indicate that the hypocotyl hooks produce a substance or substances needed in the greening process, which are translocated to the cotyledons. Indoleacetic acid, abscisic acid, gibberellin A(3), 6-benzylamino purine and delta-aminolevulinic acid do not show any activity; on the other hand, ethylene appears to replace partially the hypocotyl hooks.

Journal Article↗
PubMed1971

The effect of auxin on stress relaxation in isolated Avena coleoptiles.

In order to characterize further the mechanical properties of coleoptile cell walls, stress relaxation measurements were made on methanol-boiled sections of Avena coleoptiles. Relaxation was measured both in mechanically conditioned specimens and in specimens which had not been previously extended. In both cases the relaxation was proportional to log time. Mechanical conditioning increased the relaxation modules and decreased the relative rate of relaxation. In contrast, pretreatment of the live coleoptiles with indoleacetic acid reduced the relaxation modulus and the absolute rate of relaxation but did not affect the relative rate of relaxation. Essentially similar pictures of the mechanical properties of coleoptile walls are obtained from stress relaxation and creep tests; the wall behaves as a nonlinear viscoelastic material.

Journal Article↗