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.     

Changes in abscisic acid and indole-3-acetic acid in axillary buds of Elytrigia repens released from apical dominance

Growth of axillary buds on the rhizomes of Elytrigia repens (L) Nevski is strongly dominated by the rhizome apex, by mechanisms which may involve endogenous hormones. We determined the distribution of indole-3-acetic acid (IAA) and abscisic acid (ABA) in rhizomes and measured (by gas-chromatography-mass spectrometry) their content in axillary buds after rhizomes were decapitated. The same measurements were also made in buds induced to sprout by removing their subtending scale leaves. The ABA content tended to be higher in the apical bud and in the axillary buds than in the adjacent internodes, and tended to decline basipetally in the internodes and scale leaves. IAA was similary distributed, except that there was less difference between the buds and other rhizome parts. After rhizomes were decapitated, the ABA content of the first axillary bud declined to 20% of that of control values within 24 h, while the IAA content showed no marked tendency to change. The ABA content also declined within 12 h in the first axillary bud after rhizomes were denuded, while the content of IAA tended to increase after 6 h. These changes occurred before the length of the first axillary bud increased 24–48 h after rhizomes were decapitated or denuded. We conclude that the release of axillary buds from apical dominance in E. repens does not require IAA content to be reduced, but is associated with reduced ABA content.     

野生毛葡萄远缘杂交后代离体培养研究

以MS、1/2MS、GS三种培养基为基本培养基,以及在1/2MS培养基上分别附加6-BA 1.0mg/L+IAA 0.1mg/L和6-BA 1.0mg/L+NAA 0.1mg/L两组植物生长调节剂,对野生毛葡萄及其远缘杂交种后代213、741、196、296、B₂等6个品种(种类)进行茎段腋芽萌发离体培养试验。结果表明:1/2MS培养基为基本培养基最适宜茎段腋芽的诱导萌发;NAA对毛葡萄及其杂交种后代的萌芽生长影响明显,若单独使用,对绝大多数品种的茎段腋芽萌发有明显的抑制作用,与IAA交替使用,则对茎段腋芽萌发有促进作用。     

Effect of plant growth substances on the growth of axillary buds in cultured stem segments of Phaseolus vulgaris L.

The hormonal control of axillary bud growth was investigated in cultured stem segments of Phaseolus vulgaris L. When the stem explants were excised and implanted with their apical end in a solid nutrient medium, outgrowth of the axillary buds-located at the midline of the segment-was induced. However, if indoleacetic acid (IAA) or naphthaleneacetic acid (NAA) was included in the medium, bud growth was inhibited. The exposure of the apical end to IAA also caused bud abscission and prevented the appearance of new lateral buds.In contrast to apically inserted segments, those implanted in the control medium with their basal end showed much less bud growth. In these segments, the auxin added to the medium either had no effect or caused a slight stimulation of bud growth.The IAA transport inhibitor N-1-naphthylphthalamic acid (NPA) relieved bud growth inhibition by IAA. This suggests that the effect of IAA applied at the apical end requires the transport of IAA itself rather than a second factor. With the apical end of the segment inserted into the IAA-containing medium, simultaneous basal application of IAA relieved to some extent the inhibitory effect of the apical IAA treatment. These results, together with data presented in a related article [Lim R and Tamas I (1989) Plant Growth Regul 8: 151–164], show that the polarity of IAA transport is a critical factor in the control of axillary bud growth.Of the IAA conjugates tested for their effect on axillary bud growth, indoleacetyl alanine, indoleacetic acid ethyl ester, indoleacetyl-myo-inositol and indoleacetyl glucopyranose were strongly inhibitory when they were applied to the apical end of the stem explants. There was a modest reduction of growth by indoleacetyl glycine and indoleacetyl phenylalanine. Indoleacetyl aspartic acid and indoleglyoxylic acid had no effect.In addition to IAA and its conjugates, a number of other plant growth substances also affected axillary bud growth when applied to the apical end of stem segments. Myo-inositol caused some increase in the rate of growth, but it slightly enhanced the inhibitory effect of IAA when the two substances were added together. Gibberellic acid (GA₃) caused some stimulation of bud growth when the explants were from younger, rather than older plants. The presence of abscisic acid (ABA) in the medium had no effect on axillary bud growth. Both kinetin and zeatin caused some inhibition of axillary buds from younger plants but had the opposite effect on buds from older ones. Kinetin also enhanced the inhibitory effect of IAA when the two were applied together.In conclusion, axillary buds of cultured stem segments showed great sensitivity to auxins and certain other substances. Their growth responded to polarity effects and the interaction among different substances. Therefore, the use of cultured stem segments seems to offer a convenient, sensitive and versatile test system for the study of axillary bud growth regulation.     

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.     

Rapid in vitro propagation of Aloe barbadensis Mill

Axillary bud development and adventitious bud formation was obtained with decapitated shoot explants of Aloe barbadensis Mill. Maximal bud growth and rooting of shoots was obtained on a modified medium of Murashige and Skoog supplemented with 5 M IBA. More adventitious and axillary buds developed on nutrient media supplemented with IBA than with NAA. Axillary buds but not adventitious buds developed with IAA in the medium. Morphogenesis was inhibited by 2,4-D. Kinetin, benzyladenine and thidiazuron were toxic to the explants and did not stimulate the development of axillary of adventitious buds. The optimal temperature for bud growth and development was 25°C. Axillary bud growth and the formation of adventitious buds was slowed down at 10°C and totally inhibited by 30°C. The optimal sucrose concentration was 3% with the inhibition of bud growth and development by higher sucrose levels.     

Nitrogen, Phosphorus, and Potassium Distribution in Relation to Apical Dominance in Dwarf Bean (Phaseolus vulgaris, c.v. Canadian Wonder)

The levels and distributions of nitrogen, phosphorus, and potassiumwere followed in the axillary buds and internodes of dwarf beanplants subsequent to decapitation and application of eitherlanolin or lanolin/IAA to the cut surface of the stem. Nitrogencontinued to accumulate in decapitated internodes supplied withIAA for at least 15 days, whereas decapitated internodes nottreated with auxin showed only a slight accumulation of nitrogen.The lanolin/IAA preparation also maintained correlative inhibitionof the axillary buds for at least 15 days. However, enhancedaccumulation of N, P, and K in an IAA-treated internode didnot appear to be sufficient to deprive the axillary buds ofan adequate supply of these nutrients, for approximate balancesheets showed that more total NPK was accumulated in the internodeand axillary buds, taken together, in plants treated with plainlanolin than in those treated with IAA. Furthermore, the totalN, P, and K content per unit dry weight of the apical 5 mm ofaxillary buds was higher in the inhibited buds of IAA-treatedplants than in the elongating buds of lanolin-treated plants.Nevertheless, in dwarf bean it was found that an adequate nitrogensupply to the roots favoured lateral bud growth. From theseresults it would appear that this effect of nitrogen is an indirectone, perhaps influencing the production of substances, suchas cytokinins, stimulatory to lateral bud growth.     

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.     

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.     

棉花花芽分化及部分内源激素变化规律的研究

棉花（Ｇｏｓｓｙｐｉｕｍ ｈｉｒｓｕｔｕｍ）的腋芽原基,有的将来发育成叶枝;有的将来发育成果枝。这２种不同命运的腋芽,在其刚分化的初期就表现出了不同的解剖学特征。将来发育为叶枝的腋芽,其生长锥呈圆锥形或扁圆球形,体积较小,原套层数为１－２层;而将来发育为果枝的腋芽,其生长锥为圆柱形,顶端表面平坦,体积较大,原套层数为２－３层。从子叶展平后到肉眼可见花芽（现蕾）,连续测茎尖的内源ＡＢＡ及ＩＡＡ的含量     

Response to strigolactone treatment in chrysanthemum axillary buds is influenced by auxin transport inhibition and sucrose availability

Axillary bud outgrowth is regulated by both environmental cues and internal plant hormone signaling. Central to this regulation is the balance between auxins, cytokinins, and strigolactones. Auxins are transported basipetally and inhibit the axillary bud outgrowth indirectly by either restricting auxin export from the axillary buds to the stem (canalization model) or inducing strigolactone biosynthesis and limiting cytokinin levels (second messenger model). Both models have supporting evidence and are not mutually exclusive. In this study, we used a modified split-plate bioassay to apply different plant growth regulators to isolated stem segments of chrysanthemum and measure their effect on axillary bud growth. Results showed axillary bud outgrowth in the bioassay within 5 days after nodal stem excision. Treatments with apical auxin (IAA) inhibited bud outgrowth which was counteracted by treatments with basal cytokinins (TDZ, zeatin, 2-ip). Treatments with basal strigolactone (GR24) could inhibit axillary bud growth without an apical auxin treatment. GR24 inhibition of axillary buds could be counteracted with auxin transport inhibitors (TIBA and NPA). Treatments with sucrose in the medium resulted in stronger axillary bud growth, which could be inhibited with apical auxin treatment but not with basal strigolactone treatment. These observations provide support for both the canalization model and the second messenger model with, on the one hand, the influence of auxin transport on strigolactone inhibition of axillary buds and, on the other hand, the inhibition of axillary bud growth by strigolactone without an apical auxin source. The inability of GR24 to inhibit bud growth in a sucrose treatment raises an interesting question about the role of strigolactone and sucrose in axillary bud outgrowth and calls for further investigation.     

Effet du refroidissement local de la tige sur le transport du AIA exogène en provenance du bourgeon apical et sur la croissance des bourgeons axillaires

Effect of local cooling of the stem on exogenous 1AA transport from the apical bud and on the growth rate of axillary buds .
Local cooling (1.5°C) of the stem (on 2 or 4 cm) of Vicia faba L. cv. Aguadulce for 15 h stops the exogenous IAA transport from the apical bud and causes a high accumulation of tracers above and at the level of the cooled zone. This clamping effect is reversed after removal of the cooling system. If the cooling system is maintained for 48 h, about 25% of the exported radioactivity passes the cooled zone. However, the percentage IAA transmitted decreases to about 6.6% if the cooled zone is sufficiently wide (4 cm) and the temperature lowered to 0.7°C±0.3°C. Using a cooling system of 4 cm (0.7±0.3°C) partially releases the lowermost axillary buds (ax₁ and ax₂) from apical dominance. Yet, the rate of growth of the axillary buds is generally lower than the growth rate after decapitation of the plant.     

Effect of Abscisic Acid and Other Plant Hormones on Growth of Apical and Lateral Buds of Seedlings

The effect of abscisic acid (AbA) on the growth of lateral and apical buds was studied in seedlings of Pisum sativum and some other species. The hormone was applied in three different ways: 1) directly to the lateral bud on the second node of decapitated pea seedlings as 5 μI droplets in an ethanolic solution; 2) to the cut surface of decapitated seedlings: 3) to the apical bud of intact plants. AbA directly applied in amounts of 5 to 0.1 μg to the lateral bud of the second node of decapitated seedlings had a strong inhibitory effect on the bud. Application to the cut surface of seedlings decapitated about 5 mm above the second node resulted in slight inhibition of the lateral bud on the second node and in growth promotion of the bud on the first node. When AbA at 10 to 0.1 μg was applied to the apical bud of intact seedlings, the growth of this bud was inhibited but the lateral buds grew out. It is concluded that the release of the lateral buds from apícal dominance is the result of the inhibitory effect of AbA on growth of the apical bud and of low transport of AbA. This conclusion is supported by application of GA₃ and IAA, individually and each combined with AbA.     

Effect of phytoregulators and physiological characteristics of the explants on micropropagation of Maytenus ilicifolia

Micropropagated shoots of Maytenus ilicifolia Mart. were obtained from axillary buds cultured in Murashige & Skoog medium supplemented with 13.3 M 6-benzyladenine (BA). Addition of 1.1 M 1-indole-3-acetic acid (IAA) to the medium increased shoot elongation. The number of shoots formed was influenced by BA concentration, degree of juvenility of the explant, and by bud explant position on the stem. Cultures of buds taken from stem parts located close to the shoot tip yielded more callus than shoots, whereas axillary buds at distant positions from the apical bud yielded more shoots.Abbreviations BA 6-benzyladenine - IAA indole-3-acetic-acid     

Branching of annual shoots in common walnut (Juglans regia L.) as affected by bud production and indol-3-acetic acid (IAA) content

Relationship of bud production (axillary and terminally) of annual shoot (1Y) and/or the content of bud-derived indol-3-acetic acid (IAA) to branching of the 1Y was studied in common walnut (Juglans regia L.), cvs. Franquette and Lara. Cultivar-related branch architecture was determined. Lara tended to branch more densely than Franquette (53 vs. 42%). Significantly more fruiting off-spring shoots (FO) than vegetative ones (VO) grew-out per 1Y in both cultivars, whereas the ratio FO/VO of Lara exceeded that of Franquette by four times. An acrotonic branching pattern was more strongly expressed in Lara compared to Franquette. Bud-derived IAA was influenced by the cultivar (Franquette had 3.6 times more cumulative IAA along the 1Y than Lara), and by the relative position (terminal, subterminal, medial and basal) of the buds along 1Y. An opposite relationship between branching density and cumulative IAA content was established in both cultivars. At the 1Y relative position level, the opposite ratio between branching density and IAA content was clearly shown only on the basal position of the bud along 1Y in the Lara cultivar. Such an inconsistent linkage between bud production and the IAA spatial distribution along the 1Y illustrated that hormonal factors probably weakly affect the branching of Franquette and Lara. The length of the parent 1Y, the position of the buds along the 1Y-length, and the fate of the buds seemed to have a stronger influence on the bud out-growth and further development of the off-springs. In further analyses, seasonal fluctuations of the IAA, and the following activity of the buds should be investigated in order to improve the understanding of a complex branching phenomenon in walnut.     

In vitro propagation of Leucocroton havanensis Borhidi (Euphorbiaceae): A rare serpentine-endemic species of Cuba

Leucocroton havanensis Borhidi is an endemic plant species of Cuba able to hyperaccumulate nickel. In order to sustain the conservation of this species, an efficient protocol for its micropropagation, via axillary bud proliferation, is described. We placed apical segments from aseptic seedlings on basal medium supplemented with indole-3-acetic acid (IAA) and 6-benzylaminopurine (BAP) or thidiazuron (TDZ), individually or in combination. On a medium containing 0.5 mg L^?1 IAA and 1.0 mg L^?1 BAP explants (65.5%) developed axillary buds. Nevertheless, combinations of 0.5 mg L^?1 IAA with 0.1 mg L^?1 TDZ was the most effective treatment producing the highest number of buds per explant (30.3); while the control treatment, without growth regulators, produced no buds at all. Transfer of buds to medium supplemented with indole-3-butyric acid, indicated that 0.25 mg L^?1 is the amount of hormone required to generate roots on young buds (100%). In order to assess DNA variations in micropropagated plants, an Amplified Fragment Length Polymorphism analysis was performed and no genetic variation was detected. This study demonstrates that a high multiplication rate can be obtained by means of the reported protocol, and that plantlets can be readily hardened (96% survival) in a greenhouse by transplanting them on serpentine soil.     

Plant growth regulator and graft control of axillary bud formation and development in the TO-2 mutant tomato

The torosa-2 tomato mutant is characterized by a strong inhibition of release of axillary shoots, that is not under the control of the main apex and IAA. Microscopic examination indicated that about 70% of leaf axils do not have axillary buds. Of the growth regulators tested, gibberellic acid and cytokinins were able to modify the to-2 phenotype: increasing bud number (GA₃ treated) and developing shoots (both substances). Sequential application of growth regulators demonstrated that bud production was only affected by treatments given between sowing time and 32 days after germination. Grafting experiments indicated that endogenous root factors have no essential role in the lateral branching of the genotypes investigated. The control of axillary bud differentiation and the branching pattern in the to-2 appears to be dependent of a complex mechanism involving gibberellins and cytokinins.     

植物生长调节剂对黑木相思优树腋芽增殖及生根的影响

为建立黑木相思(Acacia melanoxylon)快繁技术体系,以含1个腋芽的无菌茎段为材料,研究了植物生长调节剂对其增殖和生根的影响。结果表明,6-BA 极易诱导黑木相思愈伤组织形成,但芽长势较差,不利于腋芽增殖体系的建立。而生长素既能诱导黑木相思生根,又能诱导腋芽增殖;将无菌茎段接入MS+IAA 0.5 mg L^-1+IBA 0.5 mg L^-1培养基中培养20 d的生根率为98.41%,培养40 d的腋芽增殖倍数为2.36,单株繁殖系数为6.57。这是首次成功建立高效、简便的生根和增殖同步发生的黑木相思直接器官发生途径的组培技术体系。     

Acropetal disappearance of PsAD1 protein in pea axillary buds after the release of apical dominance

We recently isolated PsAD1 cDNA from pea (Pisum sativum L. cv. Alaska) seedlings, whose mRNA abundantly accumulated in dormant axillary buds and disappeared after decapitation [Madoka and Mori (2000) Plant Cell Physiol. 41: 274]. To further elucidate the function of PsAD1, we investigated the temporal and spatial distribution patterns of PsAD1 protein using Western blot and immunocytochemical analyses. Western blot analyses showed that accumulation patterns of PsAD1 protein in axillary buds after decapitation and in response to IAA and 6-benzyladenine were the same as those of PsAD1 mRNA. Immunocytochemical analyses showed that (1) PsAD1 proteins were localized in the procambia, leaf primordia, apical meristem, and secondary axillary buds in the dormant axillary bud, and this distribution was the same as that of PsAD1 mRNA, (2) PsAD1 proteins acropetally disappeared after decapitation, and (3) the growth of axillary buds occurred in the same manner. These acropetal changes occur in a manner similar to the way in which the procambium differentiates into vascular tissue. These results suggest that PsAD1 plays some role in the inhibition of growth and differentiation, or in the maintenance of the dormant state in axillary buds.     

Axillary bud flowering after apical decapitation in Pharbitis in relation to photoinduction

The flowering response of axillary buds of seedlings of Pharbitis nil Choisy, cv. Violet, was examined in relation to the timing of apical bud removal (plumule including the first leaf or second leaf) before or after a flower-inductive 16-h dark period. When the apical bud was removed well before the dark period, flower buds formed on the axillary shoots that subsequently developed, but when removed just before, or after, the dark period, different results were observed depending on the timing of the apical bud removal and plant age. In the case of 8-day-old seedlings, fewer flower buds formed on the axillary shoots developing from the cotyledonary node when plumules were removed 20 to 0 h before the dark period. When the apical bud was removed after the dark period, no flower buds formed. Using 14-day-old seedlings a similar reduction of flowering response was observed on the axillary shoots developing from the first leaf node when the apical bud was removed just after the dark period. To further elucidate the relationship between apical dominance and flowering, kinetin or IAA was applied to axillary buds or the cut site where the apical bud was located. Both chemicals influenced flowering, probably by modulating apical dominance which normally forces axillary buds to be dormant.