The Different Components of the Movement and the Areas of Retention of Labelled Molecules after Application of [3H]- indolyl-acetic acid to the Apical Bud of Vicia faba

After application of [³H]-auxin (0.8 nmol) to a young leaf of Vicia faba L. cv. Aguadulce. about 6% (1.1 × 10^-2 nmol) of applied IAA enters the stem during the first 6 h of transport. This corresponds to a [³H]-auxin flux which is probably not very different from the endogenous flux. A wave of [³H]- auxin moves down to the roots mainly among preferential pathways situated in the vascular bundle. This movement is accompanied and followed by certain events: (I) In the upper part of the stem, some radioactive molecules leave the pathways of polar transport and enter the young leaves near the donor leaf. (2) In other parts of the stem, the auxin transport is highly polar. As the peak of the wave approaches and passes a node with an axillary bud. and for a few hours afterwards, there is no clearly detectable radioactivity in this bud, although the nodal tissues are very radioactive. (3) A retention of labelled molecules often occurs in the nodes. (4) Retention of label is regularly seen in the basal part of the first internode and in the hypocotyl, which together form that part of the axis where Ifle highly inhibited cotyledonary buds are found. This retention is still manifest a week after the downward transport of [³H]-auxin. (5) After 48 h. a high proportion (about 45%) of the [³H]-auxin exported by the donor leaf is found in the roots. (6) Subsequently, a part of the label returns to the upper parts of the plant, and especially to the leaves, where it normally appears to be immobilized. (7) As time goes on some labelled molecules, probably coming from different areas, enter the axillary buds.     

Abscisic acid and apical dominance in Phaseolus coccineus L.

Abscisic acid (ABA) in lanolin, applied to the internode of decapitated runner bean plants enhances the outgrowth of lateral buds. The optimum concentration of the paste is 10^-5 M. The effect of ABA is counteracted by indoleacetic acid (IAA) but not by gibberellic acid (GA₃). There is no effect when ABA is applied to the apical bud or lateral buds of intact plants. However, 13.2 ng given to the lateral buds of decapitated plants stimulate their growth, whereas higher concentrations are inhibitory. Consequently, ABA enhances growth of lateral buds directly, but only when apical dominance is already weakened. The growth of the decapitated 2^nd internode was not affected by ABA. Radioactivity from [2-¹⁴C] ABA, applied to nonelongating 2^nd internode stumps of decapitated runner bean plants moves to the lateral buds, whereas [1-¹⁴C]IAA-and [³H]GA₁-translocation is much weaker. ABA transport is inhibited if IAA or [³H]GA₁ is applied simultaneously. In elongating internodes [¹⁴C]ABA is almost completely immobile. [¹⁴C]IAA-and [³H]GA₁-translocation is not affected by ABA. The amount of radioactivity from labelled ABA, translocated to the lateral buds, is highest during the early stages of bud outgrowth.Abbreviations ABA 2,4-cis, trans-(+)-abscisic acid - GA gibberellic acid - IAA indoleacetic acid - p.l. plain lanolin     

Effect of auxin on the elongation of lateral buds and32P accumulation in 2-leaf decapitated pea seedlings

The effect of short and long term auxin treatments on the elongation of axillary buds and³²P accumulation were studied in 2-leaf decapitated Alaska pea sailings. It was found that (1) auxin delayed the elongation of the lateral buds, (2) none of the auxin concentration applied completely inhibited the elongation of axillary buds, (3) auxin had no retarding effect on the growing buds, (4) strong polarization of 32P occurred in the parts above the treated leaf, when auxin was applied for a short period just after decapitation, (5) long term auxin treatments did not induce any such polarization of 32P to the parts above the treated leaf, (6) the root acted as an alternate accumulating organ for 32P when the apex was removed and the buds were inhibited, and (7) in decapitated plants the growing buds polarized 32P.     

Changes after Decapitation in Concentrations of Indole-3-Acetic Acid and Abscisic Acid in the Larger Axillary Bud of Phaseolus vulgaris L. cv Tender Green

Early changes in the concentrations of indole-3-acetic acid (IAA) and abscisic acid (ABA) were investigated in the larger axillary bud of 2-week-old Phaseolus vulgaris L. cv Tender Green seedlings after removal of the dominant apical bud. Concentrations of these two hormones were measured at 4, 6, 8, 12 and 24 hours following decapitation of the apical bud and its subtending shoot. Quantitations were accomplished using either gas chromatography-mass spectrometry-selected ion monitoring (GS-MS-SIM) with [¹³C₆]-IAA or [²H₆]-ABA as quantitative internal standards, or by an indirect enzyme-linked immunosorbent assay, validated by GC-MS-SIM. Within 4 hours after decapitation the IAA concentration in the axillary bud had increased fivefold, remaining relatively constant thereafter. The concentration of ABA in axillary buds of decapitated plants was 30 to 70% lower than for buds of intact plants from 4 to 24 hours following decapitation. Fresh weight of buds on decapitated plants had increased by 8 hours after decapitation and this increase was even more prominent by 24 hours. Anatomical assessment of the larger axillary buds at 0, 8, and 24 hours following decapitation showed that most of the growth was due to cell expansion, especially in the intermodal region. Thus, IAA concentration in the axillary bud increases appreciably within a very few hours of decapitation. Coincidental with the rise in IAA concentration is a modest, but significant reduction in ABA concentration in these axillary buds after decapitation.     

Transport and metabolism of xylem cytokinins during lateral bud release in decapitated chickpea (Cicer arietinum) seedlings

Although cytokinins (CKs) are widely thought to have a role in promoting shoot branching, there is little data supporting a causative or even a correlative relationship between endogenous CKs and timing of bud outgrowth. We previously showed that lateral bud CK content increased rapidly following shoot decapitation. However, it is not known whether roots are the source of this CK. Here, we have used shoot decapitation to instantaneously induce lateral bud release in chickpea seedlings. This treatment rapidly alters rate and direction of solvent and solute (including CK) trafficking, which may be a passive signalling mechanism central to initiation of lateral bud release. To evaluate changes in xylem transport, intact and decapitated plants were infiltrated with [³H]zeatin riboside ([³H]ZR), a water‐soluble blue dye or [³H]H₂O by injection into the hypocotyl. All three tracers were recovered in virtually all parts of the shoot within 1 h of injection. In intact plants, solute accumulation in the lateral bud at node 1 was significantly less than in the adjacent stipule and nodal tissue. In decapitated plants, accumulation of [³H]ZR and of blue dye in the same bud position was increased 3‐ to 10‐fold relative to intact plants, whereas content of [³H]H₂O was greatly reduced indicating an increased solvent throughput. The stipule and cut stem, predicted to have high evapotranspiration rates, also showed increased solute content accompanied by enhanced depletion of [³H]H₂O. To assess whether metabolism modifies quantities of active CK reaching the buds, we followed the metabolic fate of [³H]ZR injected at physiological concentrations. Within 1 h, 80–95% of [³H]ZR was converted to other active CKs (mainly zeatin riboside‐5′phosphate (ZRMP) and zeatin (Z)), other significant, but unconfirmed metabolites some of which may be active (O‐acetylZR, O‐acetylZRMP and a compound correlated with sites of high CK‐concentrations) and inactive catabolites (adenosine, adenine, 5′AMP and water). Despite rapid metabolic degradation, the total active label, which was indicative of CK concentration in buds, increased rapidly following decapitation. It can be inferred that xylem sap CKs represent one source of active CKs appearing in lateral buds after shoot decapitation.     

Correlative inhibition of lateral bud growth in Phaseolus vulgaris L. timing of bud growth following decapitation

Summary On intact, 3-week-old plants of Phaseolus the larger bud in the axils of the primary leaves shows slow, continuous elongation growth. Release from correlative inhibition can be detected within 30 min following decapitation. When 0.1% indoleacetic acid in lanolin is applied to the decapitated stem stump, the lateral bud shows slow growth during the first 7 h, then stops completely for a further 15 h but after 2 days a further gradual increase in length is observed.The movement of ¹⁴C-labelled assimilates from the subtending primary leaf into the lateral bud increases following removal of the shoot apex. When indole acetic acid is applied to decapitated plants the ability of the buds to import ¹⁴C increases for 5–7 h and then declines to a negligible amount. Little or no radioactivity from tritiated indoleacetic acid is transported into the lateral buds of decapitated plants during the first 48 h following removal of the apex and it appears that rapid metabolism of the compound occurs in the stem tissues.     

Lateral Bud Growth in Phaseolus vulgaris L. and the Levels of Ethylene in the Bud and Adjacent Tissue

Ethephon and the ethylene inhibitors Ag⁺ and aminoethoxyvinylglycine (AVG) inhibited outgrowth of the axillary bud of thefirst trifoliate leaf in decapitated plants of Phaseolus vulgaris.Endogenous ethylene levels decreased in the stem upon decapitationalthough it is not conclusive that a causal relationship existsbetween this decrease and the release of axillary buds frominhibition. The proposition that auxin-induced ethylene is responsiblefor the suppression of axillary bud growth in the decapitatedplant when the apical shoot is replaced by auxin is not borneout in this study. Application of IAA directly to the axillarybud of intact plants gave rise to a transient increase in budgrowth. This growth increment was annulled when AVG was suppliedwith IAA to the bud despite the fact that the dosage of AVGused did not affect the normal slow growth rate of the bud ofthe intact plant or bud outgrowth resulting from shoot decapitation.     

Accumulation of 32P and [14C]sucrose in decapitated and intact shoots of the fern Davallia trichomanoides blume

The transport and accumulation of ³²P and [¹⁴C] sucrose in decapitated and intact shoot segments of the fern Davallia were studied. The apical buds of intact shoots and the expanding buds of shoots decapitated 4 weeks before application are major sinks for these nutrients. Decapitation results in a shift of ¹⁴C accumulation from the apex to the lateral buds within 36 h. This shift can be reversed in shoots decapitated for 12 h by replacing the apex. Increased ¹⁴C accumulation into the stump region occurs when decapitated shoot segments are treated with indole-3-acetic acid, and decreased label accumulation into the apical region results when intact shoots are treated with 2,3,5-triiodobenzoic acid.     

Transport of benzyl-8-14C-adenine in pea seedlings in relation to stem apical dominance

In intact, decapitated and decapitated indole-3-acetic acid (IAA) treated pea seedlings the translocation of benzyl-8-^l4C-adenin (¹⁴C-BA) from the roots was studied with regard to the release of lateral buds from apex-induced inhibition. In intact plants (controls) a substantial part of the activity was found in the apical part of the epicotyl. Decapitation resulted in the initiation of growth of lateral buds. As early as 24 h after decapitation and application of¹⁴C-BA a significantly higher activity was found in growing lateral buds (cotylars) of decapitated plants than in inhibited ones of intact or IAA-treated decapitated plants. The accumulation of¹⁴C-activity in stump tops of decapitated plants treated with IAA was associated with the thickening growth.     

Effect of decapitation on absorption,translocation, and phytotoxicity of imazamethabenz in wild oat (Avena fatua L.)

The release of apical dominance by the physical destruction in situ of the apical meristem and associated leaf primordia (decapitation) promoted the growth of tillers in non-herbicide-treated wild oat plants, as indicated by increased tiller lengths and fresh weights. At 96 h after [¹⁴C] herbicide treatment following decapitation, the absorption of [¹⁴C]imazamethabenz and total translocation of radioactivity were respectively increased by 28% and 49%. By 96 h after [¹⁴C]imazamethabenz application, the radioactivity detected in the roots of decapitated plants was 45% higher than that in the roots of nondecapitated plants while the radioactivity in tillers of decapitated plants was 2.6-fold that in tillers of intact plants. Decapitation together with foliar spraying of imazamethabenz at 200 g ha^–1 further reduced tiller fresh weight, greatly decreased the total tiller number, and thereafter significantly increased overall phytotoxicity by 32% as measured by total shoot fresh weight. The results of this study support the hypothesis that main shoot apical dominance limits translocation of applied imazamethabenz to lateral shoots, rendering tillers less susceptible to growth inhibition by the herbicide.     

Tiller Bud Suppression in Reproductive Plants of Lolium multiflorum Lam. Cv. Westerwoldicum

The control of tiller bud growth during reproductive developmentwas investigated in experimental plants ofLolium multiflorumLam. cv. Westerwoldicum that were reduced to a main axis havinga developing but unemerged ear, elongating stem internodes,a series of expanded leaves, slow-growing tiller buds and aroot system. Isolation of the ear by excision of its base, ordecapitation so as to remove the ear together with the upperleaves, promoted the movement of ¹⁴C-assimilates to tiller buds,decapitation being the more effective treatment. Applicationof 0.1 per cent indol–3yl-acetic acid (IAA) to cut tissuesof decapitated plants diverted ¹⁴C-assimilates to upper internodesbut did not reduce import by buds, whereas application of 1.0per cent IAA both diverted labelled assimilates to upper internodesand reduced bud import. Radioactivity from [¹⁴C] IAA appliedto the upper leaves or to the ear base was recovered from budsin very small amounts; larger amounts were recovered from budsfollowing the application of labelled IAA to an elongating internode,especially from the bud at the base of the treated internode.It is suggested that tiller bud suppression may be influencedby the movement of inhibitory levels of auxin into buds fromnearby elongating stem internodes, whose activity in turn maybe controlled by the developing inflorescence and upper leaves.     

Auxin transport and the regulation of petiole abscission in cotton (Gossypium hirsutum L.): A comparison of indol-3yl-acetic acid and phenylacetic acid

Distal applications of indol-3yl-acetic acid (IAA) to debladed cotyledonary petioles of cotton (Gossypium hirsutum L.) seedlings greatly delayed petiole abscission, but similar applications of phenylacetic acid (PAA) slightly accelerated abscission compared with untreated controls. Both compounds prevented abscission for at least 91 h when applied directly to the abscission zone at the base of the petiole. The contrasting effects of distal IAA and PAA on abscission were correlated with their polar transport behaviour-[1-¹⁴C]IAA underwent typical polar (basipetal) transport through isolated 30 mm petiole segments, but only a weak diffusive movement of [1-¹⁴C]PAA occurred.Removal of the shoot tip substantially delayed abscission of subtending debladed cotyledonary petioles. The promotive effect of the shoot tip on petiole abscission could be replaced in decapitated shoots by applications of either IAA or PAA to the cut surface of the stem. Following the application of [1-¹⁴C]IAA or [1-¹⁴C]PAA to the cut surface of decapitated shoots, only IAA was transported basipetally through the stem. Proximal applications of either compound stimulated the acropetal transport of [¹⁴C]sucrose applied to a subtending intact cotyledonary leaf and caused label to accumulate at the shoot tip. However, PAA was considerably less active than IAA in this response.It is concluded that whilst the inhibition of petiole abscission by distal auxin is mediated by effects of auxin in cells of the abscission zone itself, the promotion of abscission by the shoot tip (or by proximal exogenous auxin) is a remote effect which does not require basipetal auxin transport to the abscission zone. Possible mechanisms to explain this indirect effect of proximal auxin on abscission are discussed.     

Competitive canalization of PIN-dependent auxin flow from axillary buds controls pea bud outgrowth

Shoot branching is one of the major determinants of plant architecture. Polar auxin transport in stems is necessary for the control of bud outgrowth by a dominant apex. Here, we show that following decapitation in pea (Pisum sativum L.), the axillary buds establish directional auxin export by subcellular polarization of PIN auxin transporters. Apical auxin application on the decapitated stem prevents this PIN polarization and canalization of laterally applied auxin. These results support a model in which the apical and lateral auxin sources compete for primary channels of auxin transport in the stem to control the outgrowth of axillary buds.     

Transport of Benzyladenine and Gibberellin A(1) from Roots in Relation to the Dominance between the Axillary Buds of Pea (Pisum sativum L.) Cotyledons

In etiolated, 5-day-old pea (Pisum sativum L.) seedlings a significantly more intensive growth of buds situated in the axil of the excised cotyledons was observed as early as 4 hours after decapitation and excision of one cotyledon of each pair. If [8-¹⁴C]benzyladenine ([¹⁴C]BA) was applied to roots of intact plants 10 hours prior to such decapitation and excision, significantly higher both total and specific ¹⁴C activities were observed in buds situated on the side of the excised cotyledons as early as 4 hours after decapitation and excision. Although the removal of a substantial part of the root system carried out simultaneously with decapitation and excision of one cotyledon resulted in a decrease in total ¹⁴C activity of buds, nevertheless a higher accumulation of ¹⁴C activity was maintained in buds situated on the side of excised cotyledon. If [¹⁴C]BA was applied to roots of seedlings after they were decapitated and deprived of one cotyledon, both total and specific ¹⁴C activities of buds situated on the side of excised cotyledons were significantly higher as early as the end of uptake of [¹⁴C]BA by roots, i.e. after 10 hours. On the other hand, [1,2-³H]gibberellin A₁ applied to roots of intact and/or decapitated and one-cotyledon-deprived seedlings in the same way as [¹⁴C]BA did not appear in the buds until very much later and only in negligible amounts (i.e.³H activity). This indicates that the release of buds from apical dominance represents an active and selective process which can result from the ability of buds to utilize and/or synthesize only certain growth substances within a certain time interval.     

Roles for Auxin, Cytokinin, and Strigolactone in Regulating Shoot Branching

Many processes have been described in the control of shoot branching. Apical dominance is defined as the control exerted by the shoot tip on the outgrowth of axillary buds, whereas correlative inhibition includes the suppression of growth by other growing buds or shoots. The level, signaling, and/or flow of the plant hormone auxin in stems and buds is thought to be involved in these processes. In addition, RAMOSUS (RMS) branching genes in pea (Pisum sativum) control the synthesis and perception of a long-distance inhibitory branching signal produced in the stem and roots, a strigolactone or product. Auxin treatment affects the expression of RMS genes, but it is unclear whether the RMS network can regulate branching independently of auxin. Here, we explore whether apical dominance and correlative inhibition show independent or additive effects in rms mutant plants. Bud outgrowth and branch lengths are enhanced in decapitated and stem-girdled rms mutants compared with intact control plants. This may relate to an RMS-independent induction of axillary bud outgrowth by these treatments. Correlative inhibition was also apparent in rms mutant plants, again indicating an RMS-independent component. Treatments giving reductions in RMS1 and RMS5 gene expression, auxin transport, and auxin level in the main stem were not always sufficient to promote bud outgrowth. We suggest that this may relate to a failure to induce the expression of cytokinin biosynthesis genes, which always correlated with bud outgrowth in our treatments. We present a new model that accounts for apical dominance, correlative inhibition, RMS gene action, and auxin and cytokinin and their interactions in controlling the progression of buds through different control points from dormancy to sustained growth.     

硼对吲哚乙酸在植物体内运输的影响

以绿豆为指示作物,研究缺硼对侧芽生长及³H-吲哚乙酸（IAA）在完整植株体内运输的影响.结果表明：缺硼诱导侧芽生长,导致³H-IAA移动峰靠近植株顶端,茎中³H-IAA的放射性活度也低于供硼充分的植株,说明缺硼抑制了³H-IAA在植株体内的极性运输;无论缺硼与否侧芽中均未检测到³H-IAA,所以侧芽的生长与³H-IAA在其中的积累没有关系,表明硼并不是通过调节IAA在侧芽中的积累,而是通过调节IAA在主茎的移动流调控侧芽生长;给缺硼植株供硼24 h能够恢复IAA在植株体内的极性运输能力.     

Modalités du transport et du métabolisme de l'AIA 14C ou AIA 3H en relation avec la morphogenèse des bourgeons axillaires chez le Scrophularia arguta

Modes of transport and metabolism of ¹⁴ C-IAA and ³ H-IAA in relation to morphogenesis of axillary buds in Scrophularia arguta. The main objectives of this study were to investigate the morphogenetic role of IAA on the growth and development of axillary buds. After foliar applications of radioactive IAA for 6 h on intact plants of Scrophularia arguta Sol. the characteristics of auxin transport were studied by liquid scintillation counting, thin layer chromatography and microautoradiography. The main part of the radioactivity moved at a mean rate of 7 mm/h. Over long periods of transport, the tracers accumulated at the base of the axis and in the roots. The nodes were a little richer in ³H or ¹⁴C than the internodes. This fact seemed to be correlated with the vascular organization of this part of the stem. A very weak proportion of tracers was found in axillary buds. The radioactivity was to about 50% associated with the IAA molecule; the rest corresponded essentially to indolyl-4-acetyl-l-aspartic acid and indolyl-3-aldehyde. Tracers were mainly concentrated in the phloem along the whole axis and, to a lesser extent in some of the young differentiating metaxylem vessels, and in the medullary rays. No radioactivity was found in the cambial zone and in the mature xylem, nor in the parenchymas. These results support the view of an indirect role of IAA on the axillary bud growth and morphogenesis.     

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.     

Transport of exogenous auxin in two-branched dwarf pea seedlings (Pisum sativum L.)

Dwarf pea plants bearing two cotyledonary shoots were obtained by removing the epicotyl shortly after germination, and the patterns of distribution of ¹⁴C in these plants was investigated following the application of [¹⁴C]IAA to the apex of one shoot. Basipetal transport to the root system occurred, but in none of the experiments was ¹⁴C ever detected in the unlabelled shoot even after transport periods of up to 48 h. This was true both of plants with two equal growing shoots and of plants in which one shoot had become correlatively inhibited by the other, and in the latter case applied whether the dominant or subordinate shoot was labelled. In contrast, when [¹⁴C]IAA was applied to a mature foliage leaf of one shoot transfer of ¹⁴C to the other shoot took place, although the amount transported was always low. Transport of ¹⁴C from the apex of a subordinate shoot on plants bearing one growing and one inhibited shoot was severely restricted compared with the transport from the dominant shoot apex, and in some individual plants no transport at all was detected. Removal of the dominant shoot apex rapidly restored the capacity of the subordinate shoot to transport apically-applied [¹⁴C]IAA, and at the same time led to rapid cambial development and secondary vascular differentiation in the previously inhibited shoot. Applications of 1% unlabelled IAA in lanolin to the decapitated dominant shoot maintained the inhibition of cambial development in the subordinate shoot and its reduced capacity for auxin transport. These results are discussed in relation to the polarity of auxin transport in intact plants and the mechanism of correlative inhibition.Abbreviations IAA Indol-3-yl-acetic acid - TIBA 2,3,5-triiodobenzoic acid - 2,4D 2,4-dichlorophenoxyacetic acid - IAAsp Indol-3-yl-acetyl aspartic acid     

Translocation of14C-abscisic acid from roots into the aboveground part of pea (pisum sativum L.) seedlings

The translocation of¹⁴C-ABA from roots into other parts of the plant was followed in intact and decapitated pea seedlings. In intact plants ABA from roots was translocated above all into the apical part of epicotyl. In decapitated plants the regulative ability of intact apex can be partly simulated by exogenous IAA. The growth of lateral buds occurring after decapitation was associated with an intensive flow of¹⁴C-ABA from roots into released lateral buds as late as 72 h after decapitation,i.e. in the stage of intensive elongation growth of buds.