Determination of Microgram Quantities of Stevioside from Leaves of Stevia rebaudiana Bert. by Two-Dimensional Thin Layer Chromatography

A method is described for the determination of stevioside fromleaves of Stevia rebaudiana Bert. Separation upon non-activatedsilica gel impregnated with boric acid in a two-dimensionalchromatographic system followed by clution and colorimetricassay is reported. Recuperation from chromoplates is over 95%.The observed ratio of the optical density of glucose: steviosidematches that expected theoretically so that glucose may be employedas a standard where pure stevioside is not available. Effects of the plant growth retardant, ancymidol, on the growthand morphology of the shoot system of cucumber (Cucumis sativusL. ) were investigated. Ancymidol inhibited stem elongation,reducing both number and length of internodes. Reduction inleaf area, attributable to a reduction in both cell size andnumber, was accompanied by an increase in chlorophyll per unitarea. The retardant decreased apical dominance and delayed anthesis.Gibberellic acid fully reversed ancymidol-induced inhibitionof stem elongation, internode length and production, and leafexpansion. GA_4/7 and ancymidol gave a synergistic promotionof stem elongation by increasing elongation of younger internodesand increasing internode production. Synergistic promotion ofpetiole elongation by this combination was also observed. Ancymidol,applied 7 d previously either to the shoot or root, severelyreduced the level of gibberellin-like activity in bleeding sapcollected from decapitated plants.     

Interactions of the Growth Retardants Daminozide and Piproctanyl Bromide, and Gibberellins A1, A3, A4+7, A5 and A13 in Stem Extension and Inflorescence Development in Chrysanthemum morifolium Ramat

Interactions between the growth retardants daminozide (a substitutedsuccinamic acid) or piproctanyl bromide (a quaternary ammonium,piperidinium compound), and a subsequent application of a singledose (40 µg) of either gibberellin A₁, A₃, A₄₊₇ or A₁₂,showed that, in Chrysanthemum morifolium Ramat. cv. Bright GoldenAnne, a strong interdependence exists between elongation ofthe lateral shoot and the rate of development of its terminalinflorescence. A₁, A₃, and A₄₊₇ were highly active in overcoming the restrictionson both internode extension and the rate of flower-bud developmentimposed by either retardant, suggesting that these two retardanteffects are caused by a deficiency of active gibberellins (GN).In the absence of retardant, A₁, A₃, and A₄₊₇ markedly increasedstem elongation, and flowering occurred earlier than in plantsreceiving neither retardant nor GN. A₁₃ the only 20-carbon GNtested, was much less active, while A₅ had a relatively greatereffect on the time of flowering than on shoot elongation. Thus,it is not necessarily the rate of stem extension which determinesthe rate of inflorescence development. The response to different amounts of A₁, A₃ or A₁₃ (1, 5, 10,20, or 50 µg per shoot) neither suggest that different‘threshold’ levels of a particular GN are requiredto induce increases in either stem elongation or in the rateat which inflorescences develop, nor did a change in the dosegiven lead to any consistent differential effect on these twoprocesses. Chrysanthemum morifolium Ramat., stem extension, inflorescence development, growth retardants, gibberellins     

Evidence that Daminozide, but not Two Other Growth Retardants, Modifies the Fate of Applied Gibberellin A9 in Chrysanthemum morifolium Ramat

Interactions between three growth retardants and gibberellinA₉ (GA₉) in Chrysanthemum morifolium Ramat. cv. Bright GoldenAnne grown in short days have revealed that the particular retardantemployed modifies the response to this gibberellin. When plants were given a foliar spray of daminozide (a substitutedsuccinamic acid), the effects of a subsequent application ofGA₉ to each lateral shoot, on both stem length and the rateof inflorescence development, were relatively small. However,GA₉ was highly active on plants previously sprayed with piproctanylbromide (a quaternary ammonium, piperidinium compound) or onthose given a compost drench of chlorphonium chloride (a quaternaryphosphonium retardant). In the absence of GA₉ all three retardantsreduced stem length and delayed flowering. Irrespective of the extent of the responses to various dosesof GA₉ (1–50 µg per shoot), a close relationshipwas maintained between internode extension and earliness offlowering, as in previous studies with retardants and othergibberellins. On plants which had received chlorphonium chlorideGA₃ and GA₁₃ were highly active and poorly active, respectively. The data suggest that daminozide could act as a retardant notonly by blocking the biosynthesis of gibberellins but also,wholly or partly, by restricting the metabolism or action ofendogenous gibberellins.     

Gibberellin-Dependent Root Elongation in Lactuca sativa: Recovery from Growth Retardant-Suppressed Elongation with Thickening by Low concentration of GA3

Higher concentrations of growth retardants, ancymidol and AMO-1718,were required to suppress root growth than hypocotyl growthin lettuce seedlings. Gibberellic acid (GA₃) counteracted theeffect of these growth retardants, but complete recovery ofroot growth was obtained only in a narrow range of growth retardantconcentrations. A much lower concentration of GA₃ (1 nM) wasneeded for recovery of root growth from ancymidol suppressionthan that for hypocotyl growth (100 (µM). GA₃ synergisticallypromoted root growth at moderate concentrations (10–100nM) with either ancymidol or AMO-1618. Ancymidol not only suppressed root elongation but also causedthickening of the elongation zone of the root, actions whichGA₃ completely canceled. Microscopic observation showed theseeffects were mainly due to the lateral expansion and shorteningof epidermal and cortical cells. Growth kinetics of roots recorded by a computer-regulated rhizometerindicated that the lag times of both growth suppression by ancymidoland growth recovery by GA₃ were about 4 h. -Naphthaleneacetic acid (NAA) did not counteract ancymidol suppressionof root and hypocotyl growth. These results support the concept that gibberellins play anindispensable role in root elongation at an extremely lowerconcentration than in hypocotyl elongation. (Received January 7, 1987; Accepted May 14, 1987)     

Invertase Activity, Carbohydrate Metabolism and Cell Expansion in the Stem of Phaseolus vulgaris L.

In the stem of Phaseolus vulgaris L. the specific activity ofacid invertase was highest in the most rapidly elongating internode.Activity of the enzyme was very low in internodes which hadcompleted their elongation, in young internodes before the onsetof rapid elongation, and in the apical bud. From shortly afterits emergence from the apical bud the elongation of internode3 was attributable mainly to cell expansion. Total and specificactivities of acid invertase in this internode rose to a maximumat the time of most rapid elongation and then declined. Transferof plants to complete darkness, or treatment of plants withgibberellic acid (GA₃), increased the rate of internode elongationand final internode length by stimulating cell expansion. Bothtreatments rapidly increased the total and specific activitiesof acid invertase in the responding internodes; peak activitiesof the enzyme occurred at the time of most rapid cell expansion. In light-grown plants, including those treated with GA₃, rapidcell and internode elongation and high specific activities ofacid invertase were associated with high concentrations of hexosesugar and low concentrations of sucrose. As cell growth ratesand invertase activities declined, the concentration of hexosefell and that of sucrose rose. In plants transferred to darkness,stimulated cell elongation was accompanied by a rapid decreasein hexose concentration and the disappearance of sucrose, indicatingrapid utilization of hexose. No sucrose was detected in theapical tissues of light-grown plants. The results are discussed in relation to the role of acid invertasein the provision of carbon substrates for cell growth. Key words: Cell expansion, Acid invertase, Hexose, Sucrose, Phaseolus     

Apical Dominance Control in Ipomoea nil: The Influence of the Shoot Apex, Leaves and Stem

The outgrowth of lateral buds is known to be controlled by theupper shoot tissues, which include the apex, the young leavesand the upper stem. An analysis of the influence of these plantparts on axillary bud elongation in Ipomoea nil was carriedout by various treatments on these specific tissues. A restriction of elongation in the main shoot due to eitherdecapitation or shoot inversion resulted in the release of apicaldominance A non-linear type of compensating growth relationshipwas observed between the 13 cm apical growing region of thestem and the lateral buds. It was determined by decapitation,defoliation and AgNO₃ treatments that both the 13 cm stem-growthregion and the young leaves (1–5 cm in length) had a muchgreater inhibitory influence on the outgrowth of specified lateralbuds than did the stem apex (consisting of the terminal 0.5cm of the shoot). The specified lateral buds which were analyzedfor outgrowth were located a number of nodes below the shootapex. The intervening nodes were debudded. Although the importanceof young leaves in the control of apical dominance has beenpreviously recognized, the most significant result from thepresent study with Ipomoea was the strong influence of the 13cm apical growth region of the stem on the out growth of thelateral buds. Apical dominance, Ipomoea nil L., Pharbitis nil, growth region, lateral bud outgrowth, decapitation, defoliation, shoot inversion     

Interaction of growth retardants,daylength, and gibberellins A19, A20, and A1 on shoot elongation in birch and alder

Gibberellins A₁₉, A₂₀, and A₁ were applied to seedlings of birch (Betula pubescens Ehrh.) and alder (Alnus glutinosa (L.) Gaertn.) in order to test their ability to counteract growth inhibition induced by growth retardants (ancymidol and BX-112) or short day (SD, 12 h) photoperiod. Ancymidol inhibits early and BX-112 inhibits late steps in gibberellin biosynthesis. BX-112 inhibited stem elongation in both species while ancymidol, applied as a soil drench, was effective in alder only. Growth retardants affected stem elongation mainly by inhibiting elongation of internodes. All three gibberellins were equally active when applied to seedlings treated with ancymidol; however, only GA₁ was able to counteract the growth inhibition induced by BX-112. SD-induced cessation of elongation growth in birch was counteracted by GA₁, and to some degree, by GA₂₀, while GA₁₉ was inactive. SD treatment did not induce cessation of apical growth in alder. These results are consistent with the hypothesis that of gibberellins belonging to the early C-13 hydroxylation pathway, GA₁ is the only active gibberellin for stem elongation.     

Changes in Endogenous Gibberellin and Auxin Activities during First Internode Elongation in Tulip Flower Stalk

Dark treatment during the most active period of tulip shootgrowth induced rapid elongation of the first internode. Endogenousfree-form gibberellin and diffusible auxin in the first internodeincreased while bound-form gibberellin decreased after the darktreatment. Alternating dark and light treatments at 24-h intervalscaused increases in elongation of the first internode and theamounts of free-form gibberellin and diffusible auxin in thedark but their decreases in the light. TIBA treatment at thefirst node inhibited both the elongation and the increase indiffusible auxin, but did not affect the gibberellin amount.Ancymidol application prior to the dark treatment inhibitedthe increase in both free-form gibberellin and diffusible auxin.Application of gibberellin A₃ increased both elongation of thefirst internode and the amount of diffusible auxin. It alsocaused recovery from ancymidol-mediated reduction in elongationand diffusible auxin content. Dark-induced elongation of thefirst internode was inhibited when all organs above the firstinternode were excised, but endogenous free-form gibberellinincreased and bound-form gibberellin decreased. After excision,elongation of the first internode occurred only when both GA₃and IAA were applied exogenously, or when IAA was applied withdark treatment. These results indicate that dark-induced elongationof the first internode of tulip is promoted by auxin, whichis transported from the upper organs into the first internodedue to stimulation from the dark-induced increase in free-formgibberellin. Free- and bound-form gibberellins changed complementarilywith the dark and light treatments. An interconversion systembetween the two forms in the first internode and its dependenceon light conditions are also discussed. (Received June 23, 1984; Accepted March 5, 1985)     

Gibberellin-Growth Retardant Interactions on the Growth and Flowering of Clerodendrum thomsoniae

Gibberellin-growth retardant interactions on the vegetative growth and flowering of the vine Clerodendrum thomsoniae Balf. were studied using both exogenous treatments and biologically testing the acid fraction attained from the plant extract. The growth retardant, ancymidol, greatly retarded stem elongation and markedly increased flowering under inductive environments. Gibberellin A₃ (GA₃) application to the shoot tip stimulated vine growth, prevented flowering under inductive environments, and completely overcame ancymidol-induced effects. In contrast to GA₃, treatment with GA₇ had little effect on vegetative growth but increased flowering under inductive environments. The elevated activity of gibberellin-like compounds, as determined by bioassay, were similar except for a marked increase in levels in ancymidol-treated plants grown under inductive environmental conditions. Microscopic examination of the stem tip indicated that the action of the growth regulators involved the induction of floral buds. Thus, in Clerodendrum, ancymidol appears to stimulate an unknown gibberellin(s) and simultaneously acts antagonistically with GA₃.     

Internode Length in Pisum. A New Allele at the le Locus

In Pisum sativum L. a third, more severe, allele at the internodelength locus le is identified and named le^d. Plants homozygousfor le^d possess shorter internodes and appear relatively lessresponsive to GA₂₀ than comparable le (dwarf) plants. Gene le^dmay act by reducing the 3ß-hydroxylation of GA₂₀ tothe highly active GA₁ more effectively than does gene le. Theresults indicate that le is a leaky mutant and therefore thatendogenous GA₁ influences internode elongation in dwarf (le)plants. Pisum sativum, peas, internode length, genetics, gibberellin, dwarf elongation     

Effect of gibberellic acid on starch content of soybean (Glycine max L.) and its correlation with extension growth

This investigation was undertaken to study the effect of GA₃on extension growth of Glycine max L. and on starch contentof its individual internodes at maturity. The effect on hydrolyticactivity of the extract of different internodes was also studied.GA₃ stimulates the extension growth of stem by increasing theelongation of those internodes which are either in the processof elongation or being differentiated at the time of treatment.Starch content decreases with the position of the internode(from base upwards) on the intact plant. Corresponding internodeshave minimum starch content in 100 ppm GA₃-treated plants andmaximum in the controls. Internodes which show the maximum elongationdue to GA₃ treatment, show the least starch content and alsoshow maximum hydrolytic activity during the period of elongation.It is suggested that enhanced extension growth is brought aboutby enhanced mobilization of reserve food by GA₃. (Received November 21, 1967; )     

Effects of Ancymidol (a Growth Retardant) and Triarimol (a Fungicide) on the Growth, Sterols, and Gibberellins of Phaseolus vulgaris (L.)

The effect of the two substituted pyrimidines, ancymidol (a growth retardant) and triarimol (a fungicide) on Phaseolus vulgaris was studied. Both compounds retarded shoot and root elongation as well as increases in fresh weight. Both compounds caused production of ethylene-like responses when given in high dosages or when applied shortly after germination. As growth retardation was shown to occur in the absence of net increase in sterol levels, neither ancymidol nor triarimol apparently retards growth by inhibiting sterol synthesis.     

Shoot Inversion-induced Ethylene Production in the Diageotropica Tomato Mutant

Shoot inversion was found to inhibit shoot elongation over 24h in the diageotropica (dgt) mutant tomato and its isogenicparent, VNF8, by 55% and 51%, respectively. Previous studieswith normal tomato and other species would suggest that gravitystress-induced ethylene production in the inverted shoot retardselongation. Since exposure of the diageotropic shoot of thedgt mutant tomato to ethylene restores normal upward growth,it might appear that dgt is defective in its capacity to produceethylene. That shoot inversion did stimulate some ethylene productionand that the ethylene action inhibitor, AgNO₃, largely negatedthe inhibiting effect of shoot inversion on elongation in dgtstrongly suggest that the ethylene produced in the invertedshoot was responsible for its retardation of growth. Althoughethylene production in the slower growing dgt was much lessthan that in the faster-growing VNF8, the dgt shoot was foundto be much more sensitive to ethylene. Shoot inversion, ethylene, shoot elongation, diageotropica (dgt), auxin     

Growth and Physiology of One-year old Poplar (Populus) Under Elevated Atmospheric CO2 Levels

The effects of elevated atmospheric CO₂ concentrations on theecophysiological responses (gas exchange, chlorophyll a fluorescence,Rubisco activity, leaf area development) as well as on the growthand biomass production of two poplar clones (i.e. Populus trichocarpax P. deltoides clone Beaupré and P. x euramericana cloneRobusta) were examined under open top chamber conditions. Theelevated CO₂ treatment (ambient + 350 µmol mol^-1) stimulatedabove-ground biomass of clones Robusta and Beaupré afterthe first growing season by 55 and 38%, respectively. This increasedbiomass production under elevated CO₂ was associated with asignificant increase in plant height, the latter being the resultof enhanced internode elongation rather than an increased productionof leaves or internodes. Both an increased leaf area index (LAI)and a stimulated net photosynthesis per unit leaf contributedto a significantly higher stem biomass per unit leaf area, andthus to the increased above-ground biomass production underthe elevated CO₂ concentrations in both clones. The larger LAIwas caused by a larger individual leaf size and leaf growthrate; the number of leaves was not altered by the elevated CO₂treatment. The higher net leaf photosynthesis was the resultof an increase in the photochemical (maximal chlorophyll fluorescenceFm and photochemical efficiency Fv/Fm) as well as in the biochemical(increased Rubisco activity) process capacities. No significantdifferences were found in dark respiration rate, neither betweenclones nor between treatments, but specific leaf area significantlydecreased under elevated CO₂ conditions.Copyright 1995, 1999Academic Press Biomass, chlorophyll a fluorescence, elevated CO₂, growth, Populus, poplar, photosynthesis, respiration, Rubisco     

The Cellular Pathway of Short-distance Transfer of Photosynthates and Potassium in the Elongating Stem of Phaseolus vulgaris L. Stem Anatomy, Solute Transport and Pool Sizes

Based on an uniform elongation growth pattern and cellular structure,the apical 0·5-2·5 cm elongation zone of internode2 of Phaseolus vulgaris L. seedlings was selected as an experimentalsystem to study the radial pathway of photosynthate and potassiumtransfer from the phloem. An histological examination of thephloem within the elongation zone of internode 2 showed thatboth proto- and meta- phloem sieve elements were present. Theformer were fully differentiated at the commencement of elongationand became crushed as elongation proceeded. In contrast, about50% of the final number of metaphloem sieve element-companioncell complexes differentiated during the same period. The phloemdelivered some 99% of the sucrose and 72-82% of the potassiumaccumulated by the elongation zone. Solute budgets showed that,of the photosynthates and potassium entering the elongationzone, approximately 40% were retained and 60% transferred tothe shoot apex. Thus, the elongating stem acts not only as asignificant sink for photosynthates and potassium, but alsoas an axial phloem transport system to supply the shoot apex.Within the elongation zone, the principal tissue sinks weredetermined by the cellular localisation of [¹⁴C] photosynthatesand potassium by microautoradiography and ion electron microprobeanalysis respectively. About 80% of the photosynthates and potassiumwere located outside the phloem. The cortex and pith exhibitedthe greatest accumulation for photosynthates and the pith forpotassium.Copyright 1994, 1999 Academic Press Phaseolus vulgaris, elongating stem, French bean, photosynthates, potassium, radial transfer, stem anatomy, transport     

Gibberellin Regulation of Root Growth with Change in Galactose Content of Cell Walls in Pisum sativum

Root elongation of Alaska pea seedling was suppressed by higherconcentrations of growth retardants, CCC and ancymidol, thanthose required for shoot elongation. Gibberellic acid (GA³)led to recovery of ancymidol-inhibited elongation, with theconcentration (1 nM) required for roots being lower than thatfor shoots (10 µM). Ancymidol caused swelling of corticalcells in the elongating zone of the root, while GA³ completelycanceled this. These results suggest that roots require muchless gibberellin than shoots for normal elongation growth. Growth kinetics recorded by a computer-regulated rhizometerindicated that the lag periods for growth suppression by ancymidoland growth recovery by GA³ were about 10 h and 7 h, respectively. The composition of the cell wall sugars changed remarkably alongthe root axis from the tip to the base. The arabinose contentwas highest in the tip and rapidly decreased toward the base,whereas galactose complementarily increased toward the base.The thickened zone of ancymidol-treated roots had a higher galactosecontent than GA³-treated slender roots. Other neutral sugarswere not significantly influenced by ancymidol and/or GA³. Theseresults suggest that ancymidol makes cells short and thick withgalactose-rich cell walls while GA3 keeps cells extensible andslender with galactose-poor cell walls. (Received March 3, 1987; Accepted December 4, 1987)     

Internode Length in Pisum: Variation in Response to a Daylength Extension with Incandescent Light

Lines representing a range of internode length and floweringgenotypes in Pisum sativum L. were grown in 8 h of daylightfollowed by either 16 h of darkness or incandescent light. Thestem elongation response index (RI = length in 24 h ÷length in 8 h) was least in the very short internode nana types,which are grossly deficient in gibberellins (GAs), and the verylong internode slender types, which behave as if saturated withGAs. The common tall (genotype Le) and dwarf (le) types (lepartially blocks conversion of GA₂₀ to the active form, GA₁)were all markedly responsive but the peak RI (based on the mostresponsive internode) was less in tall lines (1.79 to 2.78)than in dwarf lines (2.32 to 5.01) and the peak RI tended tooccur about three to four internodes earlier in tall than indwarf lines. The cry⁸ mutation reduced the RI. (Duplicate lengthloci La and Cry are probably concerned with GA reception.) Amongle dwarf lines, genotype La cry⁸, was generally less responsivethan La Cry, La cry^c and la Cry. Data from crosses showed thaton either an le La or le la background cry⁸ segregates had alower RI than cry⁸ segregates. On an le la background, cry⁸plants were shorter than cry^c plants, cry⁸ was partially dominantto cry⁸ and segregation was clear only in long days. On an lela background, cry^c plants were shorter than cry^c plants, cry⁸was partially dominant to cry⁸ and segregation was clear inlong or short days. The very high peak RI (5.0) of the microcryptodwarfline, L57, appeared to result, in part, from a marked foreshorteningof internodes 4 to 10 in the 8 h regime. In the 24 h regimeL57 (lm) had a fairly similar growth pattern to normal (Lm)cryptodwarf types. The peak RI tended to occur at a lower internode in early thanlate flowering lines, especially among dwarf types, and genotypeswith a day neutral flowering habit (genotype sn or dne) wereless responsive than their photoperiodic counterparts (Sn Dne). White fluorescent light, given as a daylength extension, wasmuch less effective than incandescent light at stimulating stemelongation suggesting control through the phytochrome equilibrium(P_tr/P_total). Pisum sativum, garden pea, daylength extension, flowering, genotype, gibberellin, hormone receptor, incandescent light, internode length, phytochrome, stem elongation     

Factors Influencing the Distribution of Growth Between Stem and Axillary Buds in Decapitated Bean Plants

Phaseolus multiflorus plants at three stages of developmentwere decapitated either immediately below the apical bud orlower down at a point 1 cm above the insertion of the primaryleaves. Growth regulators in lanolin were applied to the cutstem surface. IAA always inhibited axillary bud elongation anddry-matter accumulation, and enhanced internode dry weight butnot elongation. GA₃ applied below the apical bud greatly increasedinternode elongation and dry weight, but simultaneously reducedbud elongation and dry-weight increase. Application of GA₃ 1cm above the buds had no effect on bud elongation in the youngestplants, but enhanced their elongation in the two older groups.IAA always antagonized GA₃-enhancement of internode extensiongrowth, whereas its effects on GA₃-enhanced dry-matter accumulationdepended on the stage of internode development. Bud elongationwas greater in plants treated with GA₃+IAA than in plants treatedonly with IAA, except in the youngest plants decapitated immediatelybelow the apical bud, where GA₃ caused a slight increase inIAA-induced bud inhibition. GA₃ increased inhibition of buddry weight by IAA in the two youngest groups of plants, butslightly reduced it in the oldest plants. No simple compensatorygrowth relationship existed between internode and buds. It wasconcluded that, (1) auxin appears to be the principal growthhormone concerned in correlative inhibition, and (2) availabilityof gibberellin to internode and buds is of importance as a modifyingfactor in auxin-regulated apical dominance by virtue of itslocal effects on growth in the internode and in the buds.     

Mode of Action of IAA and GA3 on Root and Shoot Growth of Epiphyllous Buds of Bryophyllum tubiflorum

Darkness and GA₃ stimulate the elongation of the first internodebut inhibit the production of roots while IAA inhibits internodalelongation but promotes the production of roots on epiphyllousbuds of Bryophylhum tubiflorum. Cycloheximide inhibits both,implicating the synthesis of proteins in the growth of bothroot and shoot. Even pre-treatment of buds with cycloheximidefor 4 h inhibits rooting as well as internodal elongation whenthese are subsequently transferred to IAA or GA₃. On the otherhand, pre-treatment with IAA or GA₃ even for 8 h does not alleviatethe inhibitory effect of cycloheximide suggesting that thereis a lag period between the application of these regulatorsand the synthesis of proteins caused by them.