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.     

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

Gibberellic acid (GA) stimulated both the elongation of Avena sativa stem segments and increased synthesis of cell wall material. The effects of GA on glucose metabolism, as related to cell wall synthesis, have been investigated in order to find specific events regulated by GA. GA caused a decline in the levels of glucose, glucose 6-phosphate, and fructose 6-phosphate if exogenous sugar was not supplied to the segments, whereas the hormone caused no change in the levels of glucose 6-phosphate, fructose 6-phosphate, UDP-glucose, or the adenylate energy charge if the segments were incubated in 0.1 m glucose. No GA-induced change could be demonstrated in the activities of hexokinase, phosphoglucomutase, UDP-glucose pyrophosphorylase, or polysaccharide synthetases using UDP-glucose, UDP-galactose, UDP-xylose, and UDP-arabinose as substrates. GA stimulated the activity of GDP-glucose-dependent β-glucan synthetase by 2- to 4-fold over the control. When glucan synthetase was assayed using UDP-glucose as substrate, only β-1,3-linked glucan was synthesized in vitro, whereas with GDP-glucose, only β-1,4-linked glucan was synthesized. These results suggest that one part of the mechanism by which GA stimulates cell wall synthesis concurrently with elongation in Avena stem segments may be through a stimulation of cell wall polysaccharide synthetase activity.     

Developmental changes in the gibberellin-induced growth response in stem segments of light-grown pea genotypes

The effects of GA on stem elongation were studied using segments from one tall and three dwarf light-grown pea genotypes varying in endogenous hormone content. Stem segments were cut at two distinct ages: when the fourth internode was at about 6–13% of full expansion (early-expansion) or at 18–25% of full expansion (mid-expansion). Light microscopy and flow cytometry were used to demonstrate that GA does not induce cell division in excised pea stem segments. The growth studied here was strictly elongation. Measurement of final segment length after 48 hours and high resolution measurement of growth kinetics over 20 hours using an angular position transducer were done on segments treated with hormone solutions. Our data indicate that the action of GA on stem elongation can be classified into two distinct modes. The first, apparent in early-expansion stem segments, shows distinct growth kinetics and is independent of the endogenous IAA concentration of the segments. Quantitation of IAA by GC/MS in early-expansion segments of wild type pea incubated with gibberellin shows that an increase in IAA concentration is part of the GA response in such segments. The second mode of GA action is evinced in mid-expansion segments. Whereas there is no short term (<20 h) response to GA alone (as determined by growth kinetics), there is a long term (48 h) response whose magnitude decreases across the genotypes with decreasing endogenous hormone content. Growth responses indicate that in mid-expansion segments exogenous GA acts by enhancing IAA action but appears to be unable to augment endogenous IAA content. Contradictory reports of the response of excised stem segments to GA can be reconciled when tissue genotype and developmental stage are considered.     

Role of Cell Wall Calcium in Red Light- inhibited Elongation of Hypocotyls of Etiolated Phaseolus radiatus

When the hypocotyl segments of Phaseolus radiatus L. were incubated in CaC12 (1 mmol/L) medium, the cell wall calcium was increased over threefold more than those incubated in a Ca2+ -free medium. However, red light inhibited elongation of the hypocotyl was 20% to 25% both in the medium with or without Ca2 + . The amount of calcium removed from the wall by ethylene glycol-bis (2-aminoethyl ether)-N, N, N', N'-tetraacetic acid (EGTA) ( 1 to 10 mmol/L) was 58.13 % to 75.33%, which offset the red light-inhibited elongation of the hypocotyl by 61.29% to 87.1%. Moreover, treatment with the channel blocker, verapamil ( 10 to 100 μ mol/L), wall calcium was the same as that of the darkness control, by which the red light-inhibited growth was also offset. La3 + ( 100 to 1 000 μmol/L) had no effect on wall calcium as compared to hypocotyl segments treated with red light alone, but eliminated the inhibitory effect of red light. Treatment with the calcium ionophore, A23187 (10 to 100 μmol/L), red light-inhibited elongation was abolished by 66.67% to 142.45% while wall calcium was reduced by 24.53% to 42.81%. In addition, calmodulin antagonist chlorpromazine (1 to 10 μmol/L) also counter acted the red light-induced elongation inhibition. These data indicated that exogenous Ca2+ was involved in the red light-inhibition effect, but that' did not mean that Ca2 + was not required. Perhaps Ca2 + in the wall itself was sufficient for red light-induced inhibition of hypocotyl elongation. The role of wall calcium might be quite complex, it not only acted as a signal of influx Ca2 + from the Ca2 + pool, but also played a regulatory role in the cell wall.     

Enhancement of transpiration by ethylene is responsible for absence of internodal elongation in floating rice at low humidity

Internodal elongation in floating rice (Oryza sativa) is known to be enhanced by treatment with ethylene or gibberellic acid (GA3) at high relative humidity (RH). However, ethylene-induced internodal elongation is inhibited at low RH, while GA3-induced internodal elongation is hardly affected by humidity. We examined the effects of ethylene and GA3 on the rate of transpiration in stem segments incubated at 30% or 100% RH. Ethylene promoted the transpiration of stem segments at 30% RH, but not at 100% RH, while GA3 had little effect on transpiration at either 30% or 100% RH. We propose that the absence of ethylene-induced internodal elongation at low RH is due, at least in part, to ethylene-induced transpiration.     

Effects of temperature and gibberellic acid on phospholipid composition of Avena sativa stem segments

Stem segments taken from Avena sativa plants grown at 10°, 20° or 30° varied in their phospholipid composition depending on the growth temperature; as temperature was lowered, there was a shift towards a greater proportion of unsaturated fatty acids. A significant increase was observed in the concentration of linolenic acid (18:3) as growth temperature was lowered. Although prolonged treatment of oat plants with GA₃ produced marked changes in phospholipid composition of stem segments, these changes did not always accompany the GA₃-induced growth response of segments. Treatment of stem segments with GA₃ for only 20 hr produced a significant growth response with little or no effect on phospholipid composition over this time. The data support the hypothesis that GA₃-induced growth in Avena stem segments can occur without a concomitant change in phospholipid composition.     

Genetic Dissection of the Relative Roles of Auxin and Gibberellin in the Regulation of Stem Elongation in Intact Light-Grown Peas

Exogenous gibberellin (GA) and auxin (indoleacetic acid [IAA]) strongly stimulated stem elongation in dwarf GA1-deficient le mutants of light-grown pea (Pisum sativum L.): IAA elicited a sharp increase in growth rate after 20 min followed by a slow decline; the GA response had a longer lag (3 h) and growth increased gradually with time. These responses were additive. The effect of GA was mainly in internodes less than 25% expanded, whereas that of IAA was in the older, elongating internodes. IAA stimulated growth by cell extension; GA stimulated growth by an increase in cell length and cell number. Dwarf lkb GA-response-mutant plants elongated poorly in response to GA (accounted for by an increase in cell number) but were very responsive to IAA. GA produced a substantial elongation in lkb plants only in the presence of IAA. Because lkb plants contain low levels of IAA, growth suppression in dwarf lkb mutants seems to be due to a deficiency in endogenous auxin. GA may enhance the auxin induction of cell elongation but cannot promote elongation in the absence of auxin. The effect of GA may, in part, be mediated by auxin. Auxin and GA control separate processes that together contribute to stem elongation. A deficiency in either leads to a dwarfed phenotype.     

COMPARATIVE CYTOHISTOLOGICAL STUDIES ON INHIBITION AND PROMOTION OF STEM GROWTH IN CHRYSANTHEMUM MORIFOLIUM

Sachs , R. M. (U. California, Davis), and A. M. Kopranek . Comparative cytohistological studies on inhibition and promotion of stem growth in Chrysanthemum morifolium. Amer. Jour. Bot. 50(8): 772-779. Illus. 1963.—The present study with Amo, CCC, and Phosfon,³ 3 substances which inhibit stem elongation, shows that all inhibit subapical cell expansion and division in Chrysanthemum morifolium var. ‘Indianapolis Yellow.‘ Furthermore, GA,³ in preventing the inhibition of stem elongation, maintains subapical activity at normal or greater than normal levels. For comparative purposes concentrations of the retardants and GA have been selected which completely prevent or promote the maximum rate of stem elongation. Phosfon causes complete inhibition of root growth and almost completely prevents dry matter accumulation in the tops. However, GA does not prevent such deleterious effects. Thus, GA and the growth retardants are mutually antagonistic only with respect to stem elongation and not to other aspects of growth. Furthermore, none of the retardants inhibits transverse stem growth; on the contrary transverse cell expansion and division in the subapical tissues are stimulated by the retardants, and as a result the stems of such plants are thicker than normal. GA not only prevents the thickening effect of the retardants, but, at the doses applied, GA-treated stems are considerably thinner than those of the controls, having fewer and smaller cells across the pith, cortical, and vascular tissues. Apparently, then, there is a relationship between longitudinal and transverse growth in the subapical tissues such that if one is promoted, the other is inhibited.     

Regulation of elongation growth by gibberellin in root segments of Lemna minor

Hormonal control of elongation growth was analyzed in segments excised from the elongation zone of Lemna roots. Exogenous GA3 did not promote the segment elongation but rather inhibited it. Uniconazole-P, a gibberellin biosynthesis inhibitor, significantly inhibited the segment elongation, and the inhibitory effect was completely nullified by GA3. In the epidermis, cell elongation was inhibited, but lateral cell expansion was not affected by uniconazole-P. Orientation of cortical microtubules of epidermal cells was disturbed by treatment with uniconazole-P for 12 h, and the disorganization of cortical microtubules was ameliorated by GA3. These findings suggested that disorganization of cortical microtubules induced inhibition of elongation growth of root. However, stabilization of cortical microtubules by taxol, a microtubule-stabilizing agent, did not affect the inhibition of segment elongation by uniconazole-P. These results suggested that endogenous gibberellin controls the elongation growth of root by regulating cell elongation.     

Inhibition of the synthesis of cell wall polysaccharides in oat coleoptile segments by jasmonic acid: Relevance to its growth inhibition

The inhibitory mode of action of jasmonic acid (JA) on the growth of etiolated oat (Avena sativa L. cv. Victory) coleoptile segments was studied in relation to the synthesis of cell wall polysaccharides using [¹⁴C]glucose. Exogenously applied JA significantly inhibited indoleacetic acid (IAA)-induced elongation of oat coleoptile segments and prevented the increase of the total amounts of cell wall polysaccharides in both the noncellulosic and cellulosic fractions during coleoptile growth. JA had no effect on neutral sugar compositions of hemicellulosic polysaccharides but substantially inhibited the IAA-stimulated incorporation of [¹⁴C]glucose into noncellulosic and cellulosic polysaccharides. JA-induced inhibition of growth was completely prevented by pretreating segments with 30 mm sucrose for 4 h before the addition of IAA. The endogenous levels of UDP-sugars, which are key intermediates for the synthesis of cell wall polysaccharides, were not reduced significantly by JA. Although these observations suggest that the inhibitory mode of action of JA associated with the growth of oat coleoptile segments is relevant to sugar metabolism during cell wall polysaccharide synthesis, the precise site of inhibition remains to be investigated.Abbreviations JA jasmonic acid - ABA abscisic acid - IAA indoleacetic acid - T ₀ minimum stress relaxation time - TFA trifluoroacetic acid - TCA trichloroacetic acid - HPLC high-performance liquid chromatography - EtOAc ethyl acetate - TLC thin-layer chromatography - JA-Me methyl jasmonate - GLC-SIM gas-liquid chromatography-selected ion monitoring     

TRANSLOCATION OF APPLIED GIBBERELLIN IN BEAN SEEDLINGS

The direction and extent of GA transport in Pinto beans has been studied. The increase in growth rate was used as a measure of the amount of GA which reached the stem apex. The evidence showed that a similar increase in stem growth occurred whether GA was applied to the first trifoliolate leaf or to the apex of the shoot, but considerably less elongation resulted when GA was applied to primary leaves. When leaves were treated with GA after remaining in darkness for extended times, no increase in stem elongation was observed; however, growth was promoted when the plants were returned to light. The time required for a sufficient amount of GA to be translocated from the leaf to increase stem growth is less than 1 hour. The maximum growth response was found when the treated leaf was left on the plant for 3 or more hours. A study of GA movement in two-branched plants was made. The untreated branch showed no growth response when GA was applied to the apex of the other branch, even if the dose of GA was 20 × greater than a saturating dose. Similar results were observed when GA was applied to the first trifoliolate leaf. Considerable GA moved from a mature leaf to the opposite shoot if this untreated branch had been defoliated. The pattern of GA movement to the opposite shoot was dependent on the position of the treated leaf on the shoot. It is concluded that the movement of applied GA is related to carbohydrate transport within the plant.     

IAA和GA3在调控豌豆黄化苗茎切段伸长生长中的相互作用

IAA和GA3均能促进豌豆黄化苗茎切段的伸长。IAA效应可以为GA的合成抑制剂S-3307抑制,GA3效应同样也为IAA的运输抑制剂TIBA所抑制,并且分别再施用GA3和IAA后,抑制效应又能有所解除。观察顶端切半茎切段的结果表明,IAA主要促进茎切段表皮细胞的伸长,而GA3可能主要促进内部组织细胞的伸长。观察切段横纵切片的结果则显示,IAA促进皮层细胞的伸长和增大,而GA3只促进皮层细胞的伸长。这些结果说明两者是通过不同的作用部位和方式共同调节豌豆茎切段伸长生长的。     

Inhibition of IAA-induced elongation in Avena coleoptile segments by lead: a physiological and an electron microscopic study.

A high resolution growth measuring apparatus was used to demonstrate the inhibition of auxin-induced cell elongation in oat coleoptile segments (Avena sativa L. var Holden) by lead at concentrations ranging from 2 x 10-6 M to 2 x 10-3 M. The inhibition was immediate, having no measurable lag period. Electron micrographs of lead-treated and control segments revealed that in the treated material, lead became localized as electron-dense granules in the cell walls and in vesicles associated with dictyosomes. These granules were found to be lead hydroxide phosphate by electron diffraction techniques. The possible significance of this localization and identification with regard to phosphatase activity is discussed.     

Thermoperiodic stem elongation involves transcriptional regulation of gibberellin deactivation in pea

The physiological basis of thermoperiodic stem elongation is as yet poorly understood. Thermoperiodic control of gibberellin (GA) metabolism has been suggested as an underlying mechanism. We have investigated the influence of different day and night temperature combinations on GA levels, and diurnal steady-state expression of genes involved in GA biosynthesis (LS, LH, NA, PSGA20ox1, and PsGA3ox1) and GA deactivation (PsGA2ox1 and PsGA2ox2), and related this to diurnal stem elongation in pea (Pisum sativum L. cv Torsdag). The plants were grown under a 12-h light period with an average temperature of 17 degrees C. A day temperature/night temperature combination of 13 degrees C/21 degrees C reduced stem elongation after 12 d by 30% as compared to 21 degrees C/13 degrees C. This was correlated with a 55% reduction of GA1. Although plant height correlated with GA1 content, there was no correlation between diurnal growth rhythms and GA1 content. NA, PsGA20ox1, and PsGA2ox2 showed diurnal rhythms of expression. PsGA2ox2 was up-regulated in 13 degrees C/21 degrees C (compared to 21 degrees C/13 degrees C), at certain time points, by up to 19-fold. Relative to PsGA2ox2, the expression of LS, LH, NA, PSGA20ox1, PsGA3ox1, and PsGA2ox1 was not or only slightly affected by the different temperature treatments. The sln mutant having a nonfunctional PsGA2ox1 gene product showed the same relative stem elongation response to temperature as the wild type. This supports the importance of PsGA2ox2 in mediating thermoperiodic stem elongation responses in pea. We present evidence for an important role of GA catabolism in thermoperiodic effect on stem elongation and conclude that PsGA2ox2 is the main mediator of this effect in pea.     

Gibberellins and stem growth in Arabidopsis thaliana. Effects of photoperiod on expression of the GA4 and GA5 loci.

Arabidopsis thaliana (L.) Heynh. is a quantitative long-day (LD) rosette plant in which stem growth is mediated by gibberellins (CAs). Application of GAs to plants in short-day (SD) conditions resulted in rapid stem elongation and flower formation, with GA4 and GA9 being equally effective, and GA1 showing lower activity. The effects of photoperiod on the levels of endogenous GAs were measured by combined gas chromatography-mass spectrometry with selected ion monitoring. When plants were transferred from SD to LD conditions there was a slight decrease in the level of GA53 and an increase in the levels of C19-GAs, GA9, GA20, GA1, and GA8, indicating that GA 20-oxidase activity is stimulated in LD conditions. Expression of GA5, which encodes GA 20-oxidase, was highest in elongating stems and was correlated with the rate of stem elongation. By contrast, GA4, which encodes 3 beta-hydroxylase, showed low expression in stems and its expression was not correlated with the rate of stem elongation. We conclude that stem elongation in LD conditions is at least in part due to increased expression of GA5, whereas expression of GA4 is not under photoperiodic control.     

Galactose prevents proton excretion but not IAA-induced growth in segments of azuki bean epicotyls

We investigated the effect of galactose on IAA-induced elongation and proton excretion in azuki bean (Vigna angularis Ohwi et Ohashi) segments in order to confirm whether or not protons were involved in auxin-induced growth. Galactose inhibited the IAA-induced decrease in the solution pH but had no inhibitory effect on IAA-induced growth in segments of azuki bean epicotyls. On the other hand, galactose inhibited both IAA-induced growth and proton excretion in oat (Avena sativa L.) coleoptile segments. From these results it is unlikely that IAA-induced growth is mediated by proton excretion at least in azuki bean epicotyls.Abbreviations IAA indole-3-acetic acid - FC fusicoccin     

CONTROL OF FLOWER FORMATION BY GROWTH RETARDANTS AND GIBBERELLIN IN SAMOLUS PARVIFLORUS,A LONG-DAY PLANT

The growth retardants AMO–1618 and CCC inhibited flower formation and stem elongation in Samolus parviflorus, a long-day rosette plant, under inductive conditions. The vegetative growth of the plants, as measured by leaf formation, was affected only slightly, or not affected at all. Application of gibberellic acid (GA₃) reversed completely the inhibition both of flower formation and of stem elongation caused by AMO, but relatively larger amounts of GA were required to reverse the CCC inhibition of stem elongation than that of flower formation. When applied under short-day conditions, AMO had no effect on the level of applied GA required for flower induction. When applied following long-day treatment the retardant caused some reduction of flower formation after marginal numbers of long days, but had no effect when enough long days to cause 100% flower formation were given. Other evidence indicates that the growth retardants act by inhibiting the synthesis of endogenous gibberellin. In LD plants, at least part of the action of inductive environmental conditions consists in causing an increase of gibberellin synthesis, supporting the hypothesis that relatively high GA levels are necessary for the production of the floral stimulus in this group of plants, as in long-short-day plants. The experiments with CCC indicate that stem elongation and flower formation in Samolus can be separated, and that the effect of GA on flower formation is not necessarily dependent on its effect on stem elongation.     

The role of calcium in growth induced by indole-3-acetic acid and gravity in the leaf-sheath pulvinus of oat (Avena sativa)

Leaf-sheath pulvini of excised segments from oat (Avena sativa L.) were induced to grow by treatment with 10 M indole-3-acetic acid (IAA), gravistimulation, or both, and the effects of calcium, EGTA, and calcium channel blockers on growth were evaluated. Unilaterally applied calcium (10 mM CaCl₂) significantly inhibited IAA-induced growth in upright pulvini but had no effect on growth induced by either gravity or gravity plus IAA. Calcium alone had no effect on upright pulvini. The calcium chelator EGTA alone (10 mM) stimulated growth in upright pulvini. However, EGTA had no effect on either IAA-or gravity-induced growth but slightly diminished growth in IAA-treated gravistimulated pulvini. The calcium channel blockers lanthanum chloride (25 mM), verapamil (2.5 mM), and nifedipine (2.5 mM) greatly inhibited growth as induced by IAA (50% inhibition) or IAA plus gravity (20% inhibition) but had no effect on gravistimulated pulvini. Combinations of channel blockers were similar in effect on IAA action as individual blockers. Since neither calcium ions nor EGTA significantly affected the graviresponse of pulvini, we conclude that apoplastic calcium is unimportant in leaf-sheath pulvinus gravitropism. The observation that calcium ions and calcium channel blockers inhibit IAA-induced growth, but have no effect on gravistimulated pulvini, further supports previous observations that gravistimulation alters the responsiveness of pulvini to IAA.     

The role of gibberellins in embryo axis development

The role of gibberellins (GAs) during early embryo development was examined using microspore-derived embryos (MDEs) of Brassica napus. At the globular stage of development, 10 d after initial culture (DAC) when endogenous GA(1) levels are increasing rapidly, a triazole, uniconazole, was used at 1, 33 and 100 microM to inhibit GA biosynthesis. Within this dose range there was no apparent effect of the inhibitor on embryo growth through to the early torpedo stage. However, by 25 DAC uniconazole-treated MDEs showed significantly reduced (50%) axis elongation. Addition of GA(1) at 33 microM on 14 DAC to embryos pretreated with 1 microM uniconazole on 10 DAC prevented this reduction in axis length, giving axis elongation equivalent to untreated MDEs. Application of GA(1) alone, however, did not significantly increase axis elongation. The reduced axis growth seen with uniconazole treatment was due to reduced cell elongation, but not cell number, and the co-applied GA(1) thus prevented the uniconazole-induced reduction in cell length. The elongating axis of MDEs may thus be a useful tool for examining the role of GAs in cell elongation.