Gibberellin Effect on Tryptophan Metabolism, Auxin Destruction, and Abscission in Coleus

Application of gibberellic acid (GA) to the apical region of the stem enhances ¹⁴CO₂ release from tryptophan-l-¹⁴C in cell free preparations of the apical region. Although GA when applied to the apical region markedly accelerates abscission rates of debladed petioles at the 4th node, the enhancement effect on tryptophan metabolism appears to be restricted to the apical bud region. The increased levels of diffusible auxin in Coleus stems, observed earlier by Muir and Valdovinos (1965), appear to be due to the GA effect on auxin precursor conversion rather than to an altered rate of auxin destruction. GA pre-treatment does not significantly alter destruction rates of auxin in the stem tissue. This is demonstrated by the release of ¹⁴CO₂ from IAA-1-¹⁴C by sections of internode tissue. While a multiple deblading pattern retards abscission of debladed petioles considerably, application of GA to debladed petioles at the basal region of the stem restores the normal rates of abscission at debladed distal nodes. No significant change in the abscission rates at treated nodes is observed. The GA effect on abscission at distal nodes is attributed to the effect of the growth substance on auxin precursor conversion in the apical region. In these experiments, as in the case of plants treated in the apical region with GA, auxin destruction rates in the stem are not altered significantly.     

Magnitude and Kinetics of Stem Elongation Induced by Exogenous Indole-3-Acetic Acid in Intact Light-Grown Pea Seedlings

Exogenously applied indole-3-acetic acid (IAA) strongly promoted stem elongation over the long term in intact light-grown seedlings of both dwarf (cv Progress No. 9) and tall (cv Alaska) peas (Pisum sativum L.), with the relative promotion being far greater in dwarf plants. In dwarf seedlings, solutions of IAA (between 10-4 and 10-3 M), when continuously applied to the uppermost two internodes via a cotton wick, increased whole-stem growth by at least 6-fold over the first 24 h. The magnitude of growth promotion correlated with the applied IAA concentration from 10-6 to 10-3 M, particularly over the first 6 h of application. IAA applied only to the apical bud or the uppermost internode of the seedling stimulated a biphasic growth response in the uppermost internode and the immediately lower internode, with the response in the latter being greatly delayed. This demonstrates that exogenous IAA effectively promotes growth as it is transported through intact stems. IAA withdrawal and reapplication at various times enabled the separation of the initial growth response (IGR) and prolonged growth response (PGR) induced by auxin. The IGR was inducible by at least 1 order of magnitude lower IAA concentrations than the PGR, suggesting that the process underlying the IGR is more sensitive to auxin induction. In contrast to the magnitude of the IAA effect in dwarf seedlings, applied IAA only doubled the growth in tall seedlings. These results suggest that endogenous IAA is more growth limiting in dwarf plants than in tall plants, and that auxin promotes stem elongation in the intact plant probably by the same mechanism of action as in isolated stem segments. However, since dwarf plants to which IAA was applied failed to reach the growth rate of tall plants, auxin cannot be the only limiting factor for stem growth in peas.     

Changes in Phytohormone Status in Stems and Roots after Decapitation of Pea Seedlings

Experiments were performed on the first and second internodes and 4-cm-long apical segments of main roots of pea (Pisum sativum L.) seedlings, grown in the light and decapitated above the second node on the seventh day after seed germination. Endogenous phytohormones were measured by the enzyme-linked immunosorbent assay during three days after decapitation of seedlings. The IAA level in the internodes decreased 2–3 times on the second day after decapitation of seedlings while the cytokinin level increased 5–6 times for zeatin and zeatin riboside (Z and ZR) and 1.5–2 times for isopentenyl adenine and isopentenyl adenosine (IP and IPA). In contrast to internodes, the IP and IPA contents in the roots of decapitated seedlings did not change, but the levels of Z and ZR increased 1.5–2 times compared to intact plant roots. The IAA level in the apical region of root remained almost unchanged after the removal of shoot apex. It was concluded that the apical meristem of the main root is not the site of the cytokinin response to the auxin signal coming from the stem apex and that a slight accumulation of Z and ZR after decapitation is due to upper zones of the root. There was no difference in the content of gibberellin-like substances between the internodes of intact and decapitated seedlings. However, the content of gibberellins (GA) in the root tip decreased after decapitation of seedling, which suggests an essential role of apical bud in supplying the root with GA and/or intermediates for their biosynthesis.     

Effects of the auxin-inhibiting substances raphanusanin and benzoxazolinone on apical dominance of pea seedlings

The effects of the auxin-inhibiting substances raphanusanin ((3R*,6S*)-3-[methoxy (methylthio) methyl]-2-pyrrolidinethione, raphanusanin B)and benzoxazolinone (6-methoxy-2-bezoxazolinone, MBOA) on apical dominance of pea(Pisum sativum L. cv. Alaska) seedlings were studied.Application of raphanusanin B or MBOA to the apical bud, internode, or lateralbud of pea seedlings released apical dominance in either intact orindole-3-acetic acid (IAA )-treated, decapitated plants. These results suggestthat the auxin-inhibiting substances raphanusanin B and MBOA have activity inreleasing apical dominance. Conversely, the auxin transport inhibitors2,3,4-triiodobenzoic acid (TIBA) and 1-naphthylphthalamic acid (NPA) did notstimulate lateral bud growth when they were applied directly to the lateralbud,although application to the apical bud or internode released apical dominance.Therefore, the mode of action of raphanusanin B and MBOA in apical dominance isclearly different from that of auxin transport inhibitors. Raphanusanin B andMBOA may suppress the synthesis of growth-inhibiting factor(s) of the lateralbud induced by endogenous auxin transported from the apical bud or exogenouslyapplied auxin, and/or the action of the factor(s).     

Auxin promotes gibberellin biosynthesis in decapitated tobacco plants

Excision of the apical bud (decapitation) of tobacco (Nicotiana tabacum L.) plants reduced the endogenous levels of indole-3-acetic acid (IAA), gibberellin A20 (GA20), and GA1 (the bioactive GA), in internode tissue below the excision site. Application of IAA to the stump of decapitated plants dramatically increased GA20 content, to a level 3-fold greater than in intact plants. Gibberellin A1 content was also increased by IAA. Decapitation reduced the conversion of [14C]GA19 to [14C]GA20 and of [14C]GA20 to [14C]GA1, and appeared to promote the deactivation pathway [14C]GA20 to [14C]GA29 to [14C]GA29-catabolite. Application of auxin counteracted these effects, but did not restore the conversion of [14C]GA20 to [14C]GA1 to the level found in intact plants. The results indicate that auxin is necessary for normal GA biosynthesis in stems of tobacco.     

Effect of morphactin on stem growth in relation to auxin in precooled rooted tulip bulbs

Effect of morphactin IT 3456, an auxin transport inhibitor, on tulip stem elongation induced by indole-3-acetic acid (IAA) was investigated. Tulip stem growth induced by IAA 0.1 % in lanolin paste applied on the top internode after excision of flower bud and removal of all leaves was greatly inhibited by 0.2 % morphactin IT 3456 applied on the 4th, 3rd, 2nd and 1st internode. The inhibitory effect of the morphactin on tulips stem growth promoted by IAA was restored by additional application of IAA below the morphactin treatment place. Morphactin inhibited also the growth of all internodes induced by flower bud in the absence of leaves. These results suggest a crucial role of auxin in the control growth of all internodes in tulip stem.     

Auxin transport in intact pea seedlings (Pisum sativum L.): The inhibition of transport by 2,3,5-triiodobenzoic acid

Summary The application of 2,3,5-triiodobenzoic acid (TIBA, 10 mg·g^-1 in lanolin) to the stem of intact pea seedlings (Pisum sativum L.) inhibited the basipetal transport of ¹⁴C from indoleacetic acid-1-¹⁴C (IAA-1-¹⁴C) applied to the apical bud, but not the transport of ¹⁴C in the phloem following the application of IAA-1-¹⁴C or sucrose-¹⁴C to mature foliage leaves. It was concluded that fundamentally different mechanisms of auxin transport operate in these two pathways.When TIBA was applied at the same time as, or 3.0 h after, the application of IAA-1-¹⁴C to the apical bud, ¹⁴C accumulated in the TIBA-treated and higher internodes; when TIBA was applied 24.0 h before the IAA-1-¹⁴C, transport in the stem above the TIBA-treated internode was considerably reduced. TIBA treatments did not consistently influence the total recovery of ¹⁴C, or the conversion of free IAA to indoleaspartic acid (IAAsp). These results are discussed in relation to the possible mechanism by which TIBA inhibits auxin transport,.Attention is drawn to the need for more detailed studies of the role of the phloem in the transport of endogenous auxin in the intact plant.Abbreviations TIBA 2,3,5-triiodobenzoic acid - IAAsp indoleaspartic acid     

Regulation of Auxin Levels in Coleus blumei by Ethylene

An investigation of the effects of ethylene pretreatment on several facets of auxin metabolism in Coleus blumei Benth “Scarlet Rainbow” revealed a number of changes presumably induced by the gas. Transport of indoleacetic acid-1-¹⁴C in excised segments of the uppermost internode was inhibited by about 50%. Decarboxylation of indoleacetic acid-1-¹⁴C by enzyme breis was not affected by the pretreatment. Levels of extractable native auxin in upper leaf and apical bud tissue of the pretreated plants were approximately one-half of those present in untreated plants. The rate of formation of auxin from tryptophan by enzyme breis from pretreated plants was approximately one-half that occurring in incubation mixtures containing the enzyme system from untreated plants. The conjugation of indoleacetic acid-1-¹⁴C in a form characterized chromatographically as indoleacetylaspartic acid was increased 2-fold in the upper stem region of plants pretreated with ethylene.     

Pathways of auxin transport in the intact pea seedling (Pisum sativum L.)

Summary When small colonies of the pea aphid [Acyrthosiphon pisum (Harris)] were established on the stem of Meteor Dwarf Pea seedlings (Pisum sativum L.), ¹⁴C was found in the honeydew 4.5 h after applying IAA-1-¹⁴C to a fully-expanded foliage leaf. In contrast, no activity was found in the honeydew or aphids 4.5 h after the application of IAA-1-¹⁴C to the intact apical bud even though the internode upon which the aphids were feeding contained high levels of ¹⁴C. The lack of radio-activity in aphids feeding on stems to which IAA-1-¹⁴C was applied via the apical bud was found not to be influenced by the internode position or by the transport interval allowed (up to 24 h).Radioactivity derived from either foliar or apical applications of IAA-1-¹⁴C was not transported through stem tissues killed by heat treatment. Xylem function was shown not to be impared by the heat treatment employed.It was concluded that the long-distance transport of IAA from the apical bud of intact pea seedlings does not take place in the phloem sieve tubes involved in the transport of metabolites from foliage leaves, or in the non-living tissues of the xylem.     

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.     

Enhancement of lateral bud growth in Coleus blumei BENTH. by (2-chloroethyl) trimethylammonium chloride (CCC)

The relationship of GA to apical dominance in Coleus was examinedby substituting 1 % IAA, in lanolin, for the shoot apex of CCC-treated,control and GA-treated plants containing, theoretically, hyponormal,normal and hypernormal GA levels, respectively. The greatestinhibition of lateral bud growth was obtained in the treatmentcombining 1 % IAA and 100 ppm GA, suggesting that GA may beimportant in the apical dominance of Coleus. CCC inhibited main axis growth, reduced the level of endogenousGA and caused a marked release of lateral buds from apical dominance. The significant stimulation of lateral bud growth by CCC couldnot be ascribed to reduced endogenous GA since it was not reversedby exogenous GA, or by GA plus IAA, whereas 100 ppm GA overcamethe inhibition of main axis growth by CCC. It was also shownthat the CCC stimulation was not a result of compensatory growth,that is, enhanced lateral bud growth resulting from reducedapical bud growth. The CCC effect on lateral buds was interpretedas involving a system independent of auxin and GA or else apossible immobilization of auxin in addition to inhibition ofGA biosynthesis. (Received December 5, 1967; )     

雷公藤离体快繁过程中内源激素水平的变化

对雷公藤Tripterygium wilfordii组培苗各生长发育阶段的内源激素变化进行研究。结果表明,在芽诱导过程中,培养7～14 d是芽诱导的关键性阶段,ZT含量呈上升趋势,在培养28～35 d时,ABA含量转为上升,芽体生长进入缓慢抑制阶段;在芽继代增殖过程中,GA3含量升高,ABA含量降低,GA3/IAA比值高及ABA/IAA比值低利于雷公藤芽体分化增殖;在壮苗培养过程中,IAA、GA3含量上升,但两者浓度过高或过低均不利于生长;生根培养第7 天,雷公藤根系开始生成,IAA含量达到一峰值,在生根初期,低比值ZT/IAA及高比值GA3/ABA利于雷公藤生根培养。     

Inhibition of Lettuce Seed Germination and Root Elongation by Derivatives of Auxin and Reversal by Derivatives of Cycocel

Lettuce seed germination or lettuce root elongation after germination in water was inhibited by 5.7 × 10^-6M indoleacetic acid (IAA) and designated other IAA derivatives. These IAA-inhibited growth responses were reversed by 10^-5 to 10^-6M Cycocel, (2-chloroethyl) trimethylammonium chloride, which alone was without effect. Only those Cycocel analogs, which have previously been shown to be active as plant growth retardants, were effective in reversing IAA inhibition of germination or root elongation. These results are consistent with the concept that Cycocel at low concentrations acts as an auxin antagonist. However. Cycocel did not reverse the inhibitory effects from indole-3-propionic acid or indole-3-butyric acid.     

Kinetin Transport to Axillary Buds of Dwarf Pea (Pisum sativum L.)

Decapitation resulted in the transport of significant amountsof ¹⁴C to the axillary buds from either point of application,but pretreatment of the cut internode surface of decapitatedplants with IAA (alone or in combination with unlabelled kinetin)inhibited the transport of label to the axillary buds and resultedin its accumulation in the IAA-treated region of the stem. Inintact plants to which labelled kinetin was applied to the apicalbud there was little movement of ¹⁴C beyond the internode subtendingthis bud; when labelled kinetin was applied to the roots ofintact plants, ¹⁴C accumulated in the stem and apical bud butwas not transported to the axillary buds. A considerable proportionof the applied radioactivity became incorporated into ethanol-insoluble/NaOH-solublecompounds in the apical bud of intact plants, in internodestreated with IAA, and in axillary buds released from dominanceby removal of the apical bud. The results are discussed in relation to the possible role ofhormone-directed transport of cytokinins m the regulation ofaxillary bud growth.     

Growth patterns and endogenous indole-3-acetic acid concentrations in current-year coppice shoots and seedlings of two Betula species

Apical dominance, internode elongation, radial growth and xylem cell size in coppice and apical shoots of Betula pubescens B. Pendula were determined and related to endogenous indole-3-acetic acid (IAA) levels, measured by gas chromatography-selected ion monitoring in the apical bud and at three positions along the stem. The effects of defoliation and debudding on morphological and anatomical characters and endogenous IAA levels were also investigated. The coppice shoots displayed superior stem elongation and increased branching during the initial phase of growth, after which their growth pattern was similar to that of the seedlings; however, their radial growth was greater throughout the experiment. Both plant types produced smaller-sized xylem cells at the top of the shoot than at the bottom with coppice shoots tending to form larger tracheids and smaller vessels than the seedlings. There was no consistent difference in IAA concentration between the coppice shoots and the seedlings. Defoliation and debudding reduced the IAA level in the stem within 36 h and it was still low after 25 days. Although the extent of the IAA decrease was similar in both coppice shoots and seedlings, the treatments affected the morphological and anatomical characters differently in the two plant types. The results suggest that the observed differences between seedlings and coppice shoots were not mediated through a drastic change in IAA level.     

Growth Regulators and Wood Formation in Pinus silvestris

The formation of new xylem in the spring is preceded by bud development. In decapitated pine stem the formation of xylem is arrested until the outgrowth of interfascicular buds takes place. When indole-3yl-acetic acid (IAA) is applied to the cut surfaces of decapitated stems it induces the formation of a xylem ring on the whole length of 5-ycar old trees. Naphthaleneacetic acid (NAA) causes the formation of xylem; however, the width of the growth ring is several times broader at the point of application than at the base of the leader. Cis- and trans-cinnamic acids, coumarin, L-tryptophan, kinetin (Kin), benzylaminopurine (BAP) and gibberellic acid (GA) alone do not induce cambial divisions; however, GA and the cytokinins given jointly with IAA or NAA accelerated the basipetal stimulus which has been induced by the auxins, resulting in normal xylem formation. 2,3,5-Triiodobonzoic acid (TIBA) given jointly with IAA-induced formation of compression wood in the apical part of the stem and narrow diameter tracheids at the base. When carboxyl labelled IAA or NAA are applied to pine segments it is found that the basipetal movement of IAA is much quicker than that of NAA. GA and the cytokinins increase the rate of transport of both auxins, whereas TIBA arrests the bulk of auxin in the apical part of the stem.     

Auxin-induced assimilate translocation in the bean stem (Phaseolus vulgaris L.)

Decapitation of the fully-elongated fourth internode of Phaseolus vulgaris plants resulted in the disappearance from the internode of soluble acid invertase (EC 3.2.1.26). This loss was prevented by local applications to the internode of indol-3yl-acetic acid (IAA) and, at the point of IAA application, the specific activity of the enzyme increased by up to 3 times its initial value within 48 h of treatment. IAA applications stimulated the acropetal translocation to the internode of ¹⁴C-sucrose applied to the subtending (second) trifoliate leaf 30 h after decapitation and the start of the auxin treatment. Labelled assimilates accumulated in the IAA-treated region of the internode. Following decapitation the concentration of hexose sugars in the internode fell and that of sucrose rose substantially, but these trends were reversed by IAA treatment. However, small local accumulations of sucrose occurred at the point of auxin application where tissue concentrations of IAA were greatest (determined using [1-¹⁴C] IAA).Considerable quantities of starch were present in the ground parenchyma of the internodes at the start of the experiment but, in the absence of IAA, this was remobilised within 48 h of decapitation. IAA prevented starch loss at and below its point of application to the internode, but not from more distal tissues. Cambial proliferation, radial growth and lignification were stimulated in and below IAA-treated regions of the internode. These observations are discussed in relation to the hormonal regulation of assimilate translocation in the phloem.     

Gibberellin Induced Changes in Diffusible Auxins from Savoy Cabbage

Diffusates from apices of young plants of savoy cabbage treated with gibberellie acid (GA) and apices of control plants have been examined with respect to their content of Indole auxins. Three indole Compounds were detected and identified on the basis of their chromatographic characteristics in several systems. These compounds were: glucoubrassicin, indole-3-acetic acid (IAA) and indole-3-acetonitrile (IAN). An effect of GA on the total auxin activity of the diffusate was noted 90 hours after treatment, while an increase in stem height occurred 48 hours later. This increase in auxin effect of the entire diffusates was shown bv chromogenic development and bioassay of chromatograms of diffusates to be a result of an increase in level of the IAA content. A concomitant decrease in I the glucobrassicin content was indicated. Since GA was found to have no effect on the enzymatic conversion of tryptophan or tryptamine to IAA, it is proposed that the effect of GA is on the conversion of glucobrassicin to IAA.     

The involvement of indole-3-acetic acid in the control of stem elongation in dark- and light-grown pea (Pisum sativum) seedlings

We investigated the role of auxin on stem elongation in pea (Pisum sativum L.) grown for 10d in continuous darkness or under low-irradiance blue, red, far red and white light. The third internode of treated seedlings was peeled and the tissues (epidermis and cortex+central cylinder) were separately analyzed for the concentration of free and conjugated indole-3-acetic acid (IAA). Under red, far red and white light internode elongation was linearly related with the free IAA content of all internode tissues, suggesting that phytochrome-dependent inhibition of stem growth may be mediated by a decrease of free IAA levels in pea seedlings. The correlation between IAA and internode elongation, however, did not hold for blue light-grown seedlings. The hypothesis that the growth response under low-irradiance blue light might be correlated with the lack of phytochrome B signalling and changes in gibberellin metabolism is discussed in view of current knowledge on hormonal control of stem growth.