Evidence that cytokinin controls bud size and branch form in Norway spruce

Shoot elongation in many coniferous species is predetermined during bud formation the year before the shoot extends. This implies that formation of the primordial shoot within the bud is the primary event in annual shoot growth. Hormonal factors regulating bud formation are consequently of utmost importance. We followed the levels of the endogenous cytokinins zeatin riboside (ZR) and isopentenyladenosine (iPA) in terminal buds, whorl buds and lower lateral buds of the uppermost current-year whorl shoots of 15- to 20-year-old trees of Norway spruce [ Picea abies (L.) Karst.] from June to September. Cytokinins were isolated with affinity chromatography columns, purified by high performance liquid chromatography, and quantified by ELISA. The level of ZR was low in June but increased gradually in all buds until September. Throughout the measurement period, the ZR level was highest in terminal buds and lowest in the scattered lateral, buds, with the whorl buds intermediate. The level of iPA peaked in July and decreased later without any consistent differences among the three classes of buds. The development of different kinds of buds was followed by scanning electron microscopy. We found that bud growth was greatest during August and September. The final size of primordial shoots within the buds varied considerably and the weight of the terminal bud was three times that of the whorl buds and more than five times that of the other lateral buds.
We conclude that the increase in ZR level during the period of active bud development is indicative of the importance of cytokinin for this process. Furthermore, the positive correlation between the level of ZR and bud growth during the period of predetermination of next year's branch growth suggests that this hormone indirectly controls the form of single branches in the spruce tree.     

Relations between cytokinin level, bud development and apical control in Norway spruce, Picea abies

In conifers such as Norway spruce, the extent of shoot growth is predetermined by the size and number of embryonal organs of the buds laid down the previous year. As it is known that cytokinins have a key role in bud development a possible hypothesis is that the level of cytokinin in the buds during their formation determines their size and complexity. As a first step to test this hypothesis we compared cytokinin levels in buds of different size of annual shoots from 15- to 20-year-old trees of Picea abies (L.) Karst. Apical buds from the leaders, and from branches in lower parts of the trees, were collected in April, July and August. The difference in size of the buds and the shoots growing from them was considerable in these three positions. Extracts were purified by immunoaffinity columns, and the retained compounds were separated by high-performance liquid chromatography (HPLC). Quantification was made by enzyme-linked immunosorbent assay (ELISA), and the accuracy of this method was checked by measurements with liquid chromatography-mass spectrometry (LC-MS) and UV absorption. Zeatin riboside (ZR) was the most abundant cytokinin, but isopentenyladenosine (iPA) was also present in all samples. The large apical bud of the leader contained much higher cytokinin concentrations than the considerably smaller buds from lower positions, and during the period of secondary growth in July, similar relationships were found for annual stem tissue from different positions. The possible role of ZR as a controlling factor in bud development and apical control is discussed. Our conclusion is that the level of zeatin-type cytokinins appears to play an important role in the establishment of differences in bud size and, thereby, the architecture of the tree crown.     

Bud and crown architecture of white spruce and black spruce

Needle primordia in buds and branch lengths were assessed in the crown of a plantation-grown white spruce tree. There was a gradation in needle primordia in buds in branches within the crown. The largest number of primordia was in the terminal bud of the leading main stem shoot, with the number in first-order whorl lateral shoot terminal buds decreasing from whorl 1 to whorl 4, below which buds contained a similar small number of primordia (about one-third as many as in the terminal shoot). Previous year's shoot elongation followed a similar pattern (i.e., elongation of whorl branches was greater closer to the top of the tree and elongation in the fourth through ninth whorls was about one-third that of the main stem leader). Higher order branches within whorls had within-branch gradation in shoot elongation and number of needle primordia, with older branches having as few as 16–30 primordia in buds and 3–4 cm elongation for high-order branches on older main stem whorls. There were strong correlations between the number of primordia in branch terminal buds and branch length/diameter and bud length/diameter/volume. In both black spruce and white spruce, there were strong correlations of number of needle primordia in main stem leader terminal buds with number of needle primordia in terminal buds of first and second whorl leaders.     

Lateral bud development and shoot growth in low-pruned Morus alba as affected by stem orientation

Two phases of bud activity were identified in the new growth of one-year-old erect coppice shoots on 11-year-old low-pruned stumps of mulberry (Morus alba L. cv. Shin-ichinose) in spring, the sprouting phase in which the majority of the buds, including the basal ones, sprout and elongate, and the dominance phase (starting 4–5 weeks after sprouting) during which the upper laterals begin to assert dominance and suppress the growth of lower laterals, becoming new leading shoots. In contrast, arching before sprouting markedly inhibited buds on the under side, leading to poor shoots. By late April, the sprouts on the upper side grew readily into new erect shoots, resulting in considerable dominance over those from the lateral sides. Of these erect shoots, those located closer to the stem base grew more in May and June. The effects of arching made during the sprouting phase (late April) on bud activity and shoot lengths were generally similar to those of earlier archings before spring bud bursting. Separation of the shoots from the upper and under sides by longitudinal, horizontal splitting of the arched stems in late April did not affect the inhibited elongation of the shoots from the under side. These results suggest that in the response to arching before and in late April, the effects are related to spring bud bursting and gravimorphism. In contrast, arching during and after the dominance phase (May) had no gravimorphic effects on growth of the shoots on the upper side, although there was a stimulation of outbreak of the buds on the upper side, which remained dormant during spring bud bursting. Continuous basal applications of abscisic acid in aqueous solution inhibited bud break and shoot growth of the postdormant erect stem segments, and defoliation of the new shoots markedly. In contrast, similar applications of an ethylene-releasing compound, Ethephon, depressed shoot elongation slightly, but enhanced defoliation greatly. Gibberellic acid (GA₃) stimulated shoot elongation, but depressed leaf enlargement.     

菊苣薄层培养花芽,营养芽分化中内源激素的动态变化

菊苣（Ｃｉｃｈｏｒｉｕｍ ｉｎｔｙｂｕｓＬ．）花梗薄层细胞培养于ＭＳ附加ＮＡＡ 和ＢＡ 或ＩＡＡ 和ＢＡ 的ＭＳ培养基上有花芽或营养芽分化． 花芽分化中内源ＩＡＡ、ＤＨＺ＋ ＤＨＺＲ、ｉＰＡ 含量明显增加,而Ｚ＋ ＺＲ变化不明显．营养芽分化中内源细胞分裂素含量增加明显,而ＩＡＡ 在培养前７ ｄ 含量下降,随后有所增加,在原基形成时含量达原初水平的２／３． 可见,花芽分化比营养芽分化所需内源ＩＡＡ／ＣＴＫ 比值要高     

Hormone Levels and Apical Dominance in the Aquatic Fern Marsilea drummondii A. Br

Terminal buds and successively subjacent lateral buds of the water fern, Marsilea drummondii, were examined to determine the pattern of hormone distribution in relation to apical dominance. Quantitative levels of indole-3-acetic acid (IAA), abscisic acid (ABA), zeatin and zeatin riboside (Z and ZR), and isopentenyladenosine (iPA) were determined by a solid-phase immunoassay using polycional antihormone antibodies. Enzyme-linked immunosorbent assay was used following a one-step HPLC purification procedure to obtain the free hormones. Active shoot apices contained the most IAA and Z-type cytokinins and inhibited buds the least. No significant differences in ABA levels were found leading to the conclusion that ABA did not play any role in apical dominance. The normal precedence of the most rapid outgrowth of the youngest inhibited bud as observed previously in decapitated plants was well correlated with its very high level of iPA observed in this study. The same phenomenon was observed in the median buds but with a weaker amplitude. The presence of this storage form could indicate that a bud at its entry into quiescence eventually looses the ability to hydroxylate iPA to Z-type cytokinins when it is fully inhibited. IAA and Z + ZR are concluded to be essential for lateral bud growth.     

Changes of Endogenous Hormone Contents During Floral Bud and Vegetative Bud Differentiation in Thin Cell Layer Culture of Cichorium intybus L. Explant

The sectioned thin cell layers (TCL) of flower stalk of Cichorium intybus L. were cultured in MS medium supplemented with NAA and BA or IAA and BA where floral and vegetative buds were developed from the explant. Endogenous IAA, DHZ+DHZR, iPA increased significantly during the floral bud formation, while Z+ZR remained changed. The levels of cytokinins, DHZ +DHZR, iPA, and Z-f-ZR all increased significantly during the vegetative bud formation, however IAA level was reduced during the first 7 days of culture and increased to two-thirds of initial values on the day when the bud primordia were formed. The results suggested that the initiation of floral buds was associated with a high IAA/CTK ratio, whereas the induction of vegetative bud differentiation was related to a low IAA/CTK ratio.     

The Effect of the Apical Bud on the Growth of Lateral Buds on Subterranean Shoots

The influence of the apical bud on the growth of the lateral buds on subterranean shoots was studied in Stachys sieboldiiMig. and Helianthus rigidus(Gass.) Desv. Removing and damaging the apical parts of subterranean shoots or their treatment with 2% chlorocholine chloride shoot enhanced shoot branching. The response to light of the apical bud was invariably negative: the stolons, which came out or were extracted from the soil, grew back into the ground (negative phototropism). The response to light of lateral buds was autonomous and depended on the conditions of their initiation. The lateral buds developed in darkness manifested negative phototropism when withdrawn from the soil and exposed to the light, whereas the buds developed in the light showed positive phototropism. The author concludes that the concept of apical dominance, thoroughly studied in aboveground shoots, is also valid for subterranean shoots. However, in contrast to the former, in the latter case, the apical bud does not control the growth orientation of the lateral buds.     

Periods of organogenesis in shoots of Nothofagus dombeyi (Mirb.) oersted (Nothofagaceae)

The organogenetic cycle of main-branch shoots of Nothofagus dombeyi (Nothofagaceae) was studied. Twelve samples of 52-59 parent shoots were collected from a roadside population between September 1999 and October 2000. Variations over time in the number of nodes of terminal and axillary buds, and the length, diameter and number of leaves of shoots derived from these buds (sibling shoots) were analysed. The number of nodes of buds developed by parent shoots was compared with the number of nodes of buds developed, I year later, by sibling shoots. The length, diameter and number of leaves of sibling shoots increased from October 1999 to February 2000 in those shoots with a terminal bud. However, extension of most sibling shoots, including the first five most distal leaf primordia, ceased before February due to abscission of the shoot apex. Axillary buds located most distally on a shoot had more nodes than both terminal buds and more proximal axillary buds. The longest shoots included a preformed part and a neoformed part. The organogenetic event which initiated the neoformed organs continued until early autumn, giving rise to the following year's preformation. The absence of cataphylls in terminal buds could indicate a low intensity of shoot rest. The naked terminal bud of Nothofagus spp. could be interpreted as a structure less specialized than the scaled bud found in genera of Fagaceae and Betulaceae.     

罗汉果花芽分化过程中形态及其激素水平变化特征

采用石蜡切片和酶联免疫法(ELISA)对罗汉果雄性、雌性、两性花芽分化过程的形态和激素水平变化进行观测,为罗汉果开花调控和品种选育提供科学依据。结果表明:(1)罗汉果雄性、雌性、两性花的花芽分化过程均可分为花芽未分化期、花芽分化初期、花序分化期、萼片原基分化期、花瓣原基分化期、雄蕊原基分化期和雌蕊原基分化期7个阶段。雄蕊原基分化期前,3种花芽分化过程无明显差异,各时期形态特征均依次为:茎端呈圆锥状(花芽未分化期)→茎端经半球形变成扁平状(花芽分化初期)→距茎端5~7节位处分化出穗状花序(花序分化期)→小花原基周围形成5个萼片原基(萼片原基分化期)→萼片原基内侧形成5个花瓣原基(花瓣原基分化期)。雄蕊和雌蕊原基分化期,3种花芽分化过程存在明显差异,雄蕊原基内侧出现雌蕊原基后,雄花芽雄蕊原基继续发育成雄蕊,雌蕊原基停滞生长,退为一个小突起;雌花芽雌蕊原基继续发育成雌蕊,雄蕊原基生长缓慢,退化为小花丝;两性花芽雌蕊和雄蕊原基均继续发育,形成外观正常的雌蕊和雄蕊。(2)内源激素脱落酸(ABA)、赤霉素(GAs)和玉米素核苷(ZR)含量在3种花芽分化过程中变化规律相似,即ABA含量在花芽生理分化期降低,花芽形态分化期升高,而GAs和ZR含量则基本保持不变;吲哚乙酸(IAA)含量在3种花芽分化过程中变化存在明显差异,雌花芽IAA含量在花芽生理分化期升高,花芽形态分化期逐渐降低,而雄性和两性花芽的IAA含量则基本保持不变。ABA/GAs、ABA/IAA、ZR/IAA和ZR/GAs激素含量比值在3种花芽分化过程中变化规律相似,ABA/GAs在花芽生理分化期降低,花芽形态分化期升高,而BA/IAA、ZR/IAA和ZR/GAs则基本保持不变。研究认为,罗汉果花芽分化过程经历一个"两性期",高ABA含量和ABA/GAs比值有利于罗汉果花芽分化,IAA可能对罗汉果花性分化具有重要作用。     

Shoot inversion-induced ethylene in pharbitis nil induces the release of apical dominance by restricting shoot elongation

Shoot inversion induces outgrowth of the highest lateral bud (HLB) adjacent to the bend in the stem in Pharbitis nil. In order to determine whether or not ethylene produced by shoot inversion plays a direct role in promoting or inhibiting bud outgrowth, comparisons were made of endogenous levels of ethylene in the HLB and HLB node of plants with and without inverted shoots. That no changes were found suggests that the control of apical dominance does not involve the direct action of ethylene. This conclusion is further supported by evidence that the direct application of ethylene inhibitors or ethrel to inactive or induced lateral buds has no significant effect on bud outgrowth. The hypothesis that ethylene evolved during shoot inversion indirectly promotes the outgrowth of the highest lateral bud (HLB) by restricting terminal bud (TB) growth is found to be supported by the following observations: (1) the restriction of TB growth appears to occur before the beginning of HLB outgrowth; (2) the treatment of the inverted portion of the shoot with AgNO₃, an inhibitor of ethylene action, dramatically eliminates both the restriction of TB growth and the promotion of HLB outgrowth which usually accompany shoot inversion; and (3) the treatment of the upper shoot of an upright plant with ethrel mimics shoot inversion by retarding upper shoot growth and inducing outgrowth of the lateral bud basipetal to the treated region.     

Development of shoot inHydrangea macrophylla I. Terminal and axillary buds

The structure of shoots, in particular of winter buds, ofHydrangea macrophylla was examined. The non-flower-bearing shoot is usually composed of a lower and an upper part, between which a boundary is discernible by means of a distinctly short internode. This internode is the lowermost of the upper part, and it is usually shorter than the internodes immediately above and below, although the internodes tend to shorten successively from the proximal to the distal part of the shoot. Variations exist in the following characters among the terminal bud, the axillary bud on the lower part of the shoot and the axillary bud on the upper part: (1) length of bud; (2) character of the outermost pair of leaf primordia; (3) degree of development of secondary buds in the winter bud; and (4) the number of leaf primordia. Usually, the terminal bud contains several pairs of foliage leaf primordia with a primordial inflorescence at the terminal of the bud, but the axiallary bud contains only the primordia of foliage leaves in addition to a pair of bud scales.     

Influence of root restriction and ethylene exposure on apicaldominance of petunia (Petunia xhybrida Hort. Vilm.-Andr.)

Reducing rooting volume restricted root growth during theproduction of Petunia x hybrida'Orchid and resulted in an unfavorable increase in apicaldominance. Exposing young petunia seedlings to ethylene counteracted theeffects of root restriction. Rooting volumes of 9, 28, 58, or 160mL restricted the development of lateral shoots, therebyincreasing apical dominance compared to plants grown in 162 mLrooting volumes. Ethephon, an ethylene-producing compound, increased thedevelopment of lateral shoots of seedlings grown in rooting volumes rangingfrom 28 mL to 160 mL. At a rooting volume of 9mL, ethylene exposure was not capable of reducing the growth ofthe main shoot; apical dominance remained strong in both the control andethephon-treated plants. Because lateral shoot development was not restrictedby rooting volumes greater than 160 mL, exposing these plants toethylene did not result in supplementary lateral shoot development. Levels ofindole-3-acetic acid (IAA), isopentenyladenosine (iPA), and zeatin riboside(ZR) decreased on a whole shoot basis as rooting volume decreased from 162 to58 mL. Indoleacetic acid levels in ethephon-exposed plantsdecreased 20% compared to the control. The cytokinins iPA and ZR showedno response to ethylene exposure; however, the ratio of auxin/cytokinindecreased 24% compared to the control. The decrease in theauxin/cytokinin ratio was associated with an increase in the number and lengthof lateral shoots.     

棉花花芽分化时期茎尖内源激素的变化

实验结果表明,从子叶展平后到肉眼可花芽（现蕾）,所测几种激素（ABA、IAA、GA3、iPA、ZR）的含量均表现出明显的动态变化,而且在花芽分化临界期表现出最显著的变化（出现高峰或出现低峰）。推测所测几种激素均与花芽分化有密切关系。其中ABA、GA3和CTK（iPA、ZR）在花芽分化临界期时,其含量变化均呈现出一个高峰;而IAA则在花芽分化临界期时出现一个低峰。经比较分析得知,随着花芽分化的进行,ABA/IAA、GA3／IAA、CTK／IAA均表现一个较明显的变化规律。即从子叶展平时起,其比值开始上升,到花芽开始分化时达到一个峰值,之后逐渐下降,并维持在一个较稳定的水平。显然,ABA／IAA、GA3／IAA、CTK／IAA在棉花的花芽分化过程中起着重要的调控作用。由此推测,增加植物体内的ABA、GA3、CTK的含量或降低IAA的含量,都可以促进棉花的花芽分化;反之则抑制棉花的花芽分化。     

Dormancy and spring development of lateral buds in mulberry (Morus alba)

Patterns of spring development of lateral buds of mulberry (Morus alba L. cv. Shin-ichinose) coppice shoots on 11-year-old low-pruned stumps varied in response to girdling, pruning and arching. The erect controls showed a weak acrotonic (apex-favoring) growth habit, in which the majority of the buds, including the basal ones, sprouted and elongated in mid- and late April, and hence there was a prolonged imposition of dominance on the upper laterals in mid- and late May. In contrast, early spring girdling or pruning enhanced the activity of the upper buds of the proximal (lower) halves of the girdled stems or of the pruned stems, resulting in considerable dominance of the laterals from such buds in late April. Arching markedly inhibited buds on the under side of the arched stems, leading to poor shoots. By late April, the buds on the adaxial (upper) side readily grew into new vertical shoots, which dominated over the lateral ones. When studied by a multiple-node-cutting test, increased length of segments of post-dormant mulberry stems was accompanied by decreased bud activity of the segments and by decreased breaking ability of the lower buds within the segments, suggesting the importance of roots in the weak acrotonic habit of the erect stem in spring. By contrast, the acropetal influences of the attached stems can in part affect dominance relationships, perhaps mediated through competition for factors translocated from the roots. Continuous basal applications of abscisic acid inhibited bud break and shoot growth of the postdormant stem segments, but these inhibitory effects could be reversed by applied gibberellic acid A₃ (GA₃). Two phases of lateral bud dormancy in erect mulberry coppice shoots were identified. The first was characterized by a smaller breaking capacity in the upper buds than in the lower ones and hence by a basitonic (base-favoring) gradient in bud growth potential. The second phase corresponded to a restoration of these capabilities in the upper buds and to a change towards a linear gradient in bud growth potential, with disappearance of the dormant condition, in February and March. This gradient change during dormancy release may represent the physiological basis for the weak acrotonic habit of erect mulberry stems in spring.     

香荚兰花芽分化至萌发期内源激素的变化

以香荚兰 (Vanillafragrans)为材料 ,研究不同栽培条件下花芽分化和萌发期内源激素变化 ,分析和探讨内源激素在花芽分化和萌发中的作用 ,香荚兰花芽分化时期茎里的激素含量降低 ,芽里激素含量升高 ,其中相对高的ZR和ZR ABA有利于分化 ,IAA和IAA ABA的一定增加也利于分化 ,过高或没有IAA的增加则不利于花芽分化。大多数花芽形成于倒垂茎蔓上 ,花芽分化期 (11～ 12月 ) ,倒垂茎蔓的茎里生长类激素含量降低大于竖立茎蔓 ,芽的激素含量增高则多于竖立茎蔓 ,倒垂茎蔓的这种变化可能是有利于花芽分化。香荚兰生长中顶端优势明显 ,去顶后侧芽里ZR、GA、IAA增高 ,这与 11～ 12月去顶促进倒垂茎蔓开花可能有关。     

Apple shoot architecture: evidence for strong variability of bud size and composition and hydraulics within a branching zone

* In the apple tree (Malus domestica), shoot architecture - the distribution of lateral bud types and growth along the parent shoot - has been extensively investigated. The distal zone of a shoot is characterized by a high proportion of vegetative or floral axillary branches mixed with latent buds and aborted laterals. The hypothesis tested here was that bud development was related to hydraulic conductance of the sap pathway to the bud, independently of an acrotonic (proximal vs distal) effect. * The distal zone of 1-yr-old shoots was studied on five cultivars for bud size and composition (number of appendages) and hydraulic conductance before bud burst. * Bud size, composition and hydraulic conductance were highly variable for all cultivars. A positive correlation was demonstrated between both the number of cataphylls and green-leaf primordia, and hydraulic conductance. Cultivar and bud size affected the intercept of these relationships more than the slope, suggesting similar scaling between these variables, but different hydraulic efficiencies. A great proportion of small buds were also characterized by null values of hydraulic conductance. * This study suggests that hydraulically mediated competition exists between adjacent buds within the same branching zone, prefiguring the variability of lateral types in the following growing season. It is hypothesized that this developmental patterning is driven by hydraulic characteristics of the whole metamer, including the subtending leaf, during bud development.     

Ontogeny of proventitious epicormic buds in Quercus petraea. I. In the 5 years following initiation

 The persistence of large epicormic shoots is one of the main factors that reduces timber quality and value in Quercus petraea. The early phases of epicormic shoot formation, i.e. the initiation of the epicormic buds, their survival and their proliferation over the years, are not clearly understood. In the present work, we studied the initiation of the axillary buds giving rise to epicormic buds and shoots, and followed their behaviour during the first 5 years using both scanning electron microscopy and light microscopy. Two types of proventitious epicormic buds have been identified. The first type has small axillary buds associated with the rings of bud-scale scars which are found at the base and tip of each growth unit. These buds are made of a terminal meristem surrounded only by scales; no leaf primordium is detected. During the second and third years of epicormic life, meristematic areas appear in the scale axil. Progressively, the meristematic areas organize into secondary bud primordia composed solely of the terminal meristem surrounded by scales. The second type of epicormic bud has secondary buds produced by a large axillary bud when this large bud either developed into a shoot or partially abscised. The epicormic potential in Q. petraea is characterized by a balance between the epicormic buds in apparent rest, enclosing meristematic areas and secondary bud primordia, and their mortality over the years. Received: 22 January 1998 / Accepted: 8 May 1998     

Preformed and neoformed extension of shoots and sylleptic branching in relation to shoot length in Tsuga canadensis

Summary Shoot systems developed over 3 successive years were investigated on 55 understorey Tsuga canadensis (L.) Carr. trees. Paired comparisons of preformed-leaf content of terminal buds and numbers of leaves produced on new shoots showed that neoformed leaves were produced in large numbers. Parent-shoot character was not useful in predicting numbers of preformed leaves, was better related to total leaves produced, but left the majority of the variation unexplained. This reflected the capacity of any terminal bud to produce a shoot with more or less neoformation, depending on conditions for growth. All shoots over 6 cm long produced sylleptic shoots that bore from two to many leaves and were arranged in a mesitonic pattern along the parent. Some of the longer sylleptic shoots produced lateral buds or second-order sylleptic shoots. Monopodial second-year extensions of sylleptic-shoot axes followed an acrotonic pattern, as did proleptic shoots from the few lateral buds borne on the parent shoots. Such lateral buds were more frequent on shorter parent shoots: they typically occurred near the proximal and distal ends. Duration of shoot extension was positively correlated with shoot length: terminal buds became evident as shoot extension neared cessation.