Effects of gibberellic Acid and sucrose on the growth of oat (Avena) stem segments

Gibberellic acid induced growth in Avena (oat) stem segments within 35 minutes after hormone application. The total elongation elicited by gibberellic acid was greater than 15 times the control growth. The sensitivity of the segments to low concentrations of gibberellic acid (1 pmole) and the specificity of the segments to the gibberellin class of hormones suggest that oat stem segments would be a valuable tool for gibberellin bioassays. Both gibberellic acid-induced growth and control growth are temperature-dependent and showed a Q₁₀ of two or greater. Although the most apparent effect of gibberellic acid was to promote the uptake of water into the internode, the hormone also promoted transport of endogenous substrate and the uptake of exogenous substrate into the growing region. The growth promotion was accomplished without an apparent increase in osmotic pressure.     

Regulation of cell wall synthesis in Avena stem segments by gibberellic Acid

Gibberellic acid induces (a) increased elongation of Avena sativa stem segments, (b) increased formation of cell wall material, measured on the basis of dry weight, and (c) increased incorporation of ¹⁴C-glucose into all fractions of the cell wall material. This increased incorporation of radioactivity correlates well with increased formation of cell wall material and shows a time-course pattern similar to the time course of the elongation response. Approximately one hour after the application of gibberellic acid, the rates both of growth and of incorporation of radioactivity accelerate to about 2-fold over the control rate. Gibberellic acid does not stimulate the incorporation of labeled glucose into the cell wall material simply by increasing the rate of uptake of glucose by internodal cells. The stimulation of the incorporation of ¹⁴C-glucose into cell wall material, which reflects the stimulation of cell wall synthesis, seems to be an important and relatively early effect of gibberellic acid in this system and probably contributes significantly to the elongation response elicited by the hormone.     

Regulation of invertase levels in Avena stem segments by gibberellic Acid, sucrose, glucose, and fructose

Gibberellic acid and sucrose play significant roles in the increases in invertase and growth in Avena stem segments. About 80% of invertase is readily solubilized, whereas the rest is in the cell wall fraction. The levels of both types of invertase change in a similar manner in the response to gibberellic acid and sucrose treatment. The work described here was carried out with only the soluble enzyme. In response to a treatment, the level of invertase activity typically follows a pattern of increase followed by decrease; the increase in activity is approximately correlated with the active growth phase, whereas the decrease in activity is initiated when growth of the segments slows. A continuous supply of gibberellic acid retards the decline of enzyme activity. When gibberellic acid was pulsed to the segments treated with or without sucrose, the level of invertase activity increased at least twice as high in the presence of sucrose as in its absence, but the lag period is longer with sucrose present. Cycloheximide treatments effectively abolish the gibberellic acid-promoted growth, and the level of enzyme activity drops rapidly. Decay of invertase activity in response to cycloheximide treatment occurs regardless of gibberellic acid or sucrose treatment or both, and it is generally faster when the inhibitor is administered at the peak of enzyme induction than when given at its rising phase. Pulses with sucrose, glucose, fructose, or glucose + fructose elevate the level of invertase significantly with a lag of about 5 to 10 hours. The increase in invertase activity elicited by a sucrose pulse is about one-third that caused by a gibberellic acid pulse given at a comparable time during mid-phase of enzyme induction, and the lag before the enzyme activity increases is nearly twice as long for sucrose as for gibberellic acid. Moreover, the gibberellic acid pulse results in about three times more growth than the sucrose pulse. Our studies support the view that gibberellic acid, as well as substrate (sucrose) and end products (glucose and fructose), play a significant role in regulating invertase levels in Avena stem tissue, and that such regulation provides a mechanism for increasing the level of soluble saccharides needed for gibberellic acid-promoted growth.     

Effect of gibberellic Acid on elongation and longevity of coleus petioles

The effects of gibberellic acid on the longevity and elongation of variously aged, debladed petioles of Coleus blumei were studied, with particular reference to the hypotheses 1) that auxin increases longevity by increasing growth, and 2) that gibberellic acid acts by increasing the endogenous levels of auxin.

Gibberellic acid, substituted for the leaf blades, significantly decreased longevity of younger petioles, as measured by days or hours to abscission. Gibberellic acid also decreased the longevity resulting from 0.1% indoleacetic acid. This is the opposite of the effect expected if it is increasing auxin levels in the petiole.

In its effect on elongation of younger petioles, however, gibberellic acid did act in the direction expected if it were increasing effective levels of auxin in the petiole. The elongation rate from 0.1% gibberellic acid plus 0.1% indoleacetic acid in lanolin was as large or larger than that for 1.0% indoleacetic acid.

Petioles which were 10 or more weeks old (i.e., at positions 5+ below the apical bud were not affected by 0.1% gibberellic acid in either longevity or rate of elongation, with or without 0.1% indoleacetic acid. Since 1.0% indoleacetic acid increases both longevity and elongation rate of these petioles over 0.1% indoleacetic acid, gibberellic acid is clearly not acting on older petioles as if it were increasing effective auxin levels).

     

Role of Indole-3-acetic Acid and Gibberellin in the Control of Internodal Elongation in Avena Stem Segments: Long Term Growth Kinetics

Exogenous application of indoleacetic acid results in a significant suppression of the linear growth that is promoted by exogenous gibberellic acid in Avena stem segments in a fashion similar to that previously noted in Avena leaf base segments (van Overbeek and Dowding, 1961, Fourth International Conference Plant Growth Regulation). Treatment with the auxin transport inhibitors, methyl-2-chloro-9-hydroxyfluorene-(9)-carboxylate (CFM) or 2,3,5-triiodobenzoic acid (TIBA), alone promotes elongation growth of the stem segments over that of control growth. This effect is interpreted as being due to the interference in the transport of native indoleacetic acid by CFM and TIBA, thus removing the inhibitory effect of native indoleacetic acid on gibberellin-promoted growth in the internodal intercalary meristem. This results in a greater promotion of internodal growth by native gibberellins. In the presence of (2-chloroethyl) trimethylammonium chloride (CCC), the growth-promoting effects of CFM and TIBA are decreased, and the antiauxin, PCIB (4-chloro-phenoxyisobutyric acid), has no growth-promoting effects whatsoever. These results indicate that the CFM and TIBA-promoted growth require the continuous presence of gibberellins. They further support the view that native indoleacetic acid acts as a growth suppressor hormone in its regulation of gibberellin-promoted internodal extension in Avena shoots.     

Interaction of indoleacetic Acid and gibberellic Acid in the short-term growth kinetics of oat stem segments

Gibberellins and auxins are the only hormones known that promote growth in oat (Avena sativa) stem segments, but when applied together, indoleacetic acid inhibits gibberellic acid-induced growth appreciably. This study shows that in addition to this inhibitory role, indoleacetic acid shortens the response time of the tissue to gibberellic acid.     

Synergism between Gibberellin and Auxin in the Growth of Intact Tomato

Gibberellin A_4&7 was more effective than gibberellic acid in increasing shoot elongation when applied to the apex of intact Lycopersicum esculentum seedlings of Tiny Tim, a dwarf cultivar, and Winsall, a tall cultivar. After 14 days, gibberellic acid and gibberellin A_4&7 stimulated growth of the dwarf more than the tall tomato. In tall tomato the application of indole-3-acetic acid alone (6.1 μg/plant) showed an inhibitory growth effect, but when applied with 17.5 μg per plant of gibberellic acid, it had a synergistic effect at 7 days but not at 14 days. When the auxin concentration was reduced to 0.61 μg per plant a synergistic effect was observed on tall plants at 7 and 14 days between indole-3-acetic acid and gibberellic acid. Application of gibberellin A_4&7 with auxin did not give a synergistic response in tall or dwarf tomato.     

Further biological characteristics of galactoglucomannan oligosaccharides

The biological activity of cell wall-derived galactoglucomannan oligosaccharides (GGMOs) was dependent on their chemical structure. Galactosyl side chains linked to the glucomanno-core influenced their inhibition of elongation growth of pea (Pisum sativum L. cv. Tyrkys) stem segments induced by 2,4-dichlorophenoxyacetic acid (2,4-D). Reduction of the number of galactosyl side chains in GGMOs caused stimulation of the endogenous growth. Modification on the glucomanno-reducing end did not affect significantly the activity of these oligosaccharides. GGMOs inhibited also the elongation induced by indole-3-acetic acid (IAA) and gibberellic acid (GA₃). In the presence of IAA the elongation growth was inhibited to 20 – 35 % after 24 h of incubation depending on GGMOs concentrations (1 μM, 10 nM, 0.1 nM), similarly as in the presence of 2,4-D, which confirms the hypothesis of GGMOs antiauxin properties. The elongation induced by GA₃ was inhibited to 25 – 60 %, however, the time course of inhibition was different compared with IAA and 2,4-D. The highest inhibition was determined already after 6 h of incubation with a significant decrease after this time. The results indicated a competition between GGMOs and growth regulators.     

Does Ethylene Play a Role in the Release of Lateral Buds (Tillers) from Apical Dominance in Oats?

The growth of lateral buds (tillers), which are undergoing release from apical dominance, was measured in upright and gravistimulated intact Avena sativa L. cv. `Victory' (oat) shoots as well as in isolated Avena stem segments treated with kinetin and sucrose. During release, the tiller bud initially shows a slow rate of elongation accompanied by swelling. It is followed by a more rapid rate of elongation. Ethylene (C₂H₄) production in shoot segments containing a tiller bud was found to occur at the onset of tiller swelling during gravistimulation as well as during inflorescence emergence. Exogenous application of indoleacetic acid or C₂H₄ inhibits kinetin-induced tiller bud swelling and elongation. However, stem segments pulsed for 24 hours in C₂H₄ or the C₂H₄ biosynthesis precursor, 1-amino-cyclopropane-1-carboxylic acid (ACC) and then transferred to kinetin and sucrose, showed a significant increase in swelling elongation as compared with segments maintained under the same conditions but without C₂H₄ or ACC in the pulse. Segments pulsed for 24 hours with kinetin and sucrose plus the ACC biosynthesis inhibitor, aminoethoxyvinylglycine, or the C₂H₄ action inhibitor, CO₂, then transferred to kinetin and sucrose medium, showed inhibition of tiller swelling during the pulse and of subsequent elongation. These results indicate that C₂H₄ plays a role in promoting tiller swelling during the onset of tiller release from apical dominance and may act as a modulator hormone in promoting tiller elongation in the presence of cytokinin.     

Altered Growth Response to Exogenous Auxin and Gibberellic Acid by Gravistimulation in Pulvini of Avena sativa

Pulvini of excised segments from oats (Avena sativa L. cv Victory) were treated unilaterally with indoleacetic acid (IAA) or gibberellic acid (GA₃) with or without gravistimulation to assess the effect of gravistimulation on hormone action. Optimum pulvinus elongation growth (millimeters) and segment curvature (degrees) over 24 hours were produced by 100 micromolar IAA in vertical segments. The curvature response to IAA at levels greater than 100 micromolar, applied to the lower sides of gravistimulated (90°) pulvini, was significantly less than the response to identical levels in vertical segments. Furthermore, the bending response of pulvini to 100 micromolar IAA did not vary significantly over a range of presentation angles between 0 and 90°. In contrast, the response to IAA at levels less than 10 micromolar, with gravistimulation, was approximately the sum of the responses to gravistimulation alone and to IAA without gravistimulation. This was observed over a range of presentation angles. Also, GA₃ (0.3-30 micromolar) applied to the lower sides of horizontal segments significantly enhanced pulvinus growth and segment curvature, although exogenous GA₃ over a range of concentrations had no effect on pulvinus elongation growth or segment curvature in vertical segments. The response to GA₃ (10 micromolar) plus IAA (1.0 or 100 micromolar) was additive for either vertical or horizontal segments. These results indicate that gravistimulation produces changes in pulvinus responsiveness to both IAA and GA₃ and that the changes are unique for each growth regulator. It is suggested that the changes in responsiveness may result from processes at the cellular level other than changes in hormonal sensitivity.     

Interactions between abscisic acid, ethylene and gibberellin in internodal elongation in floating rice: the promotive effect of abscisic acid at low humidity

The enhancement of internodal elongation in floating or deepwater rice (Oryza sativa L. cv. Habiganj Aman II) by treatment with ethylene or gibberellic acid (GA₃) at high relative humidity (RH) is inhibited by abscisic acid (ABA). Here, we examined the interactive effects of ethylene, gibberellin (GA) and ABA at low RH on internodal elongation of deepwater rice stem segments. Although ethylene alone hardly promoted internodal elongation of stem sections at 30% RH, it enhanced the internodal elongation induced by GA₃. Application of ABA alone to stem segments had no effect on internodal elongation. However, in the presence of ethylene and GA₃ at 30% RH, ABA further promoted internodal elongation. This promotive effect of ABA was not found in the internodes of stem segments treated either with ethylene or with GA₃ at 30% RH or in the internodes of stem segments treated with ethylene and/or GA₃ at 100% RH.     

Red Light-inhibited Mesocotyl Elongation in Maize Seedlings: I. The Auxin Hypothesis

Red light-inhibited mesocotyl elongation, which occurs in intact Zea mays L. seedlings, was studied in excised segments which included the coleoptile (or parts therefrom) and apical centimeter of the mesocotyl. Experiments took into account, first, the ability of the segments to regenerate auxin supply sites, and, second, that auxin uptake can be greatly reduced if there is no cut surface, apical to the elongating cells, to act as a port of entry. In all cases, auxin completely reversed the inhibition of elongation by light. The results support the hypothesis that light regulates mesocotyl elongation by controlling auxin supply from the coleoptile. Sucrose concentration had no effect on auxin reversal of light-inhibited elongation, but relatively high concentrations of gibberellic acid (10 μm) could substitute for auxin in this system.     

Regulation of lettuce hypocotyl elongation by gibberellic acid. Correlation between cell elongation, stress-relaxation properties of the cell wall and wall polysaccharide content

Lettuce hypocotyl elongation caused by gibberellic acid wasstrongly inhibited by coumarin and dichlobenil, known inhibitorsof cellulose biosyndiesis. Stress-relaxation analysis of thecell wall revealed that gibberellic acid induces a decreasein both minimum relaxation time (To) and relaxation rate (b)and an increase in maximum relaxation time (Tm), when gibberellicacid stimulates hypocotyl elongation. Both coumarin and dichlobenilnullified the effect of gibberellic acid on changes in To, Tmand b values. The content of pectic, hemicellulosic and cellulosic substancesin the cell wall increased per hypocotyl but decreased per unithypocotyl length, in response to gibberellic acid treatment.Particularly, gibberellic acid caused a substantial increasein cellulose content per hypocotyl but a decrease per unit length.A good correlation existed between the decrease in To and thedecrease in hemicellulose content per unit lengdi of the cellwall. The increase in Tm was correlated with the decrease incellulose content per unit length of the cell wall. The decreasein b was correlated with the decrease in the content of bothcellulose and hemicellulose per unit length. Based on these results, we discuss the role of polysaccharidemetabolism of the cell wall in gibberellic acid-induced lettucehypocotyl elongation and the nature of gibberellic acid-inducedbiochemical modifications of the cell wall, which are representedby changes in stress-relaxation properties of the cell wall. ¹Present address: Department of Anatomy, Aichi Medical University,Nagakutecho, Aichigun, Aichi 480-11, Japan. (Received September 22, 1975; )     

Modification of cell wall polysaccharides during cell elongation in Phaseolus vulgaris hypocotyls

The effect of gibberellic acid (GA) and naphthylacetic acid (NAA) on hypocotyl elongation and cell wall polysaccharides was studied using Phaseolus vulgaris seedlings grown in light condition. The hypocotyl was demarcated into two segments — one near the root was called lower and the one near the cotyledon was called upper. The upper segment showed a typical sigmoidal growth curve while lower segment did not show any growth at all. GA promoted the growth of upper segment while NAA showed clear inhibition in both the segments. Xyloglucan content showed a clear inverse correlation with growth. Pectic polysaccharides did not show a clear trend, though showed an initial inverse correlation with growth. It is concluded that degradation of low and high molecular weight xyloglucans are involved in cell wall loosening which in turn may be responsible for the elongation growth of Phaseolus hypocotyls in light.     

Inhibition of Gibberellic Acid-induced Elongation in Avena Stem Segments by a Substituted Pyrimidine

Avena stem segments, which respond with high amplitude, specificity, and sensitivity to gibberellic acid, were used to study the inhibition of gibberellin-induced elongation by the growth retardant alpha-cyclopropyl-alpha-(4-methoxyphenyl)-5-pyrimidine methanol (EL-531). It was found that EL-531 strongly inhibits gibberellic acid-induced elongation in this system at a concentration of 1 mm. From a double-reciprocal plot of elongation and gibberellic acid concentration, it seems that EL-531 and gibberellic acid do not compete reversibly for the same site of action. Also, because EL-531 effectively inhibits elongation in internodal tissue dissected away from the node and leaf sheath, it cannot be acting primarily by inhibiting the synthesis or transport of the leaf sheath factor(s). Because EL-531 causes lateral expansion of the stem segments as well as increased diameters of epidermal cells, in a manner very similar to the effects of colchicine, it is suggested that EL-531 inhibits gibberellic acid-induced elongation by somehow interfering with the orientation of the products of cell wall synthesis.     

Effect of Gibberellic Acid on Dwarf and Normal Pea Plants

Gibberellic acid at concentrations between 10 and 100 mg/1 greatly stimulated the elongation growth of intact dwarf pea plant but showed little or no effect on that of Alaska pea. It showed no effect on the elongation growth of excised stem segments of either dwarf or normal pea when given alone. Indole-3-acetic acid stimulated the elongation of excised segments of both varieties. Gibberellic acid synergistically enhanced the indole-3-acetic acid-induced elongation of excised segments. Tryptophan also stimulated the elongation of these segments. Gibberellic acid showed a synergistic effect on the tryptophan-induced elongation, as on the indole-3-acetic acidinduced one. Gibberellic acid reduced the lag period of tryptophan-induced elongation, suggesting that gibberellic acid promotes the conversion of tryptophan to auxin.     

Gibberellin Substitution for the Requirement of the Cotyledons in Stem Elongation in Pisum sativum Seedlings

The removal of the cotyledons from 8-day-old light-grown Pisum sativum cv. Alaska seedlings caused a reduction in the rate of stem elongation to 50% of the intact control value. Gibberellic acid restored the stem elongation rate of decotylized plants to the level of the intact controls. The effect of decotylization was to lower both the rate of node formation and the rate of internode elongation. The steady state rate of internode elongation was reduced to 50% of the control rate by decotylization. Applied gibberellic acid did not restore the normal rate of node formation nor the lag in internode elongation caused by decotylization, but gibberellic acid did restore the normal steady state rate of internode elongation. Analysis of variance demonstrated an interaction between the cotyledons and applied gibberellic acid. 2-Isopropyl-4-dimethylamino-5-methyl phenyl-1-piperidine carboxylate methyl chloride inhibited internode elongation to the same extent in both intact and decotylized plants. The results indicate that the cotyledons are an effective source of gibberellin for the young pea seedling.     

Promotion of elongation and acid invertase activity in Phaseolus vulgaris L. internode segments by phenylacetic acid

Phenylacetic acid (PAA) significantly stimulated the elongation of isolated Phaseolus vulgaris internodal segments and prevented the decline in acid invertase specific activity observed in segments incubated in the absence of growth substances. Unlike IAA, which stimulated both elongation and invertase activity over a very wide range of concentrations (<10^-4 - 1 mol.m^-3; optimum 10^-2 mol.m^-3), the response to PAA was restricted to a much narrower range of concentrations (3 × 10^-2 - 1 mol.m^-3; optimum ca. 1–2 × 10^-1mol.m^-3). At the optimum concentration of PAA, the stimulation of both responses was about 63–75% of that induced by the optimum concentration of IAA. The differences in the concentration range and magnitude of the responses to IAA and PAA were not due to differences in uptake of the two compounds. The stimulation of elongation by both compounds was prevented by 3.6 × 10^-2mol.m^-3 cycloheximide (CH), and acid invertase activites were greatly reduced compared with samples treated with growth substances alone. A saturating concentration of the specific auxin efflux carrier inhibitor N-1-naphthylphthalamic acid (NPA) slightly promoted the growth of control segments, probably by reducing the loss of residual endogenous auxin to the incubation medium. The elongation induced by PAA at its optimum concentration was considerably greater than the elongation induced by NPA, indicating that PAA did not cause growth by preventing the loss of endogenous auxin from the segments. Elongation responses to combinations of IAA and PAA suggested that the compounds were acting additively and that they were affecting growth by the same mechanism.     

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.     

Effect of Gibberellic Acid on the Elongation Rate of Agrostis alba Root Hairs

The influence of gibberellic acid over a wide range of concentrations on the rate of elongation of root hairs of redtop grass was investigated. The rate of root hair elongation was increased by GA over the concentration range of 10^?7 to 10^?12 M inclusive, with peak stimulation occurring at 10^?6 M. Although root hair growth was slightly accelerated by 10^?6 M GA, this concentration damaged many root hairs and caused some to stop growing altogether. Rate of root hair elongation was reduced to less than 84% of the control by 10^?5 M GA.