Content of endogenous cytokinins in intact and transversally cut leaf blades of Bryophyllum crenatum

An increased growth of marginal buds on excised leaves from vegetative plants of Bryophyllum crenatum following transversal separation of leaf blades into three segments (apical, medial and basal) as compared with non-separated leaves, was associated with increased endogeneous cytokinin-like substances. This cycokinin increase can be demonstrated seven days after the separation and before the growth of marginal bud primordia. The level of endogenous cytokinin-like substances increases in a similar direction as the formation and growth of marginal buds, from the base of leaf blade to its apex.

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Zusammenfassung Der Anstieg von Marginalknospenbildung an den von den vegetativen Pflanzen Bryophyllum crenatum isolierten Blättern, der durch eine Querteilung der Blattspreiten in drei Segmente (apikales, mittleres und basales) gegenüber den intakten Kontrollblattspreiten nachweisbar ist, ist mit dem Anstieg der endogenen Zytokininähnlichen Stoffe verknüpft. Diesen Anstieg des Zytokininspiegels in der geteilten Blattspreite, gegenüber der intakten, kann man binnen sieben Tage nach der Querteilung der Blattspreite beweisen, d.h. in der Zeit vor dem Wachstum der Marginalknospenanlagen. Der endogene Zytokininspiegel steigt dabei von der Basis zum Apex der Blattspreite an, ähnlich wie die Marginalknospenbildung.

     

The correlative effect of the stem apex on the level of endogenous gibberellins inBryophyllum crenatum leaves

Following a one-side transverse incision made into the middle stem part ofBryophyllum crenatum below one of the two opposite leaves, the leaf is exposed to an enhanced effect of correlatively inhibitory substances passing out of the apical part of the stem. Within 28 days this results in a formation of adventitious roots above the incision. The flow of correlatively inhibiting substances interrupted by the incision induces, as early as within 24 h, a decrease in the level of endogenous gibberellins in the leaf above the inoision.     

Endogenous gibberellins and auxins in the stem ofbryophyllum crenatum in relationship to its polarity

The analyses for the content of endogenous gibberollins and auxins in leaves and adjacent stem internodia of intactBryophyllum crenatum plants revealed that the level of gibberellins increases in tho direction from the apex to the stem base, whereas in the case of auxins the trend in the increase is tho reverse. This corresponds to the results of morphological experiments, according to which the apical stem part appears as affected by phytohormones of an inhibitory character, the basal part as affected by stimulatory ones.     

Polarity of the stem and ontogenetic changes in sunflow (Helianthus annuus L.) leaves in relation to endogenou cytokinins and ethylene

The content of endogenous cytokinin-like substances and the release of ethylene were determined in leaves of different insertion of sunflower plants during their ontogeny. The content of cytokinin-like substances was highest in the leaves on the middle part of the stem (that is in leaves just before full expansion), with a decrease occurring both towards the base and the apex of the stem, when followed at four growth phases (vegetative plants, plants with inflorescence diameter up to 0.5 cm, plants with inflorescence diameter up to 3 cm, and plants in flower). Changes in the content of cytokinin-like substances during the ontogeny of the leaf also corresponded to this pattern. Data obtained with the leaf at the third node from the basis of the stem showed that the level of cytokinin-like substances first sharply increased, and then after reaching maximal value (at the time when leaf blade area reached approximately 70 % of the final value) slowly and continuously decreased. The highest amount of ethylene released from the leaves was recorded in basal leaves and then also in apical leaves, whereas the leaves with the largest blade area situated at the central part of the stem released the lowest amount of ethylene. This pattern was repeatedly found at all four selected growth phases of sunflower plants.     

Ontogenetic changes in the content of endogenous gibberellins inBryophyllum crenatum leaves with respect to stem polarity

The content of endogenous gibberellins was estimated in leaves adjacent to the individual nodes ofBryophyllum crenatum in five ontogenetic periods (July 10, August 12, September 17, October 22, and December 8). Their content decreased from the stem base to the apex when 5 to 7 leaf pairs were developed (July 10 and August 12). Before transition to the generative state when 8 leaf pairs were formed (Sept. 17 and Oct. 22) the content of gibberellins was gradually increased in the apical leaves and decreased in the basal ones. This change resulted in the increasing gradient of leaf gibberellins from the stem base to the apex just before flowering (Dec. 8). The content of endogenous gibberellins increases during the leaf ontogenesis up to the beginning of its senescence. This trend occurs first in the basal leaves (I –III) where the initial increase is followed by the decline in the gibberellin content. On the other hand gibberellins in leaves derived from the apical nodes (VII–VIII) only gradually increase in the course of leaf ontogenesis.     

The release of primordia of marginal buds onbryophyllum crenatum leaves from growth inhibition in relationship to the level of endogenous IAA

The growth of primordia of marginal buds (marginals) which differentiate on leaf margins is correlatively inhibited on intactBryophyllum crenatum plants. Following leaf isolation, the marginals are released from this correlative inhibition, which process is accompanied within 2 to 10 h after leaf isolation with a decrease in the content of endogenous IAA in the leaf blade. This decrease can be enhanced by transversal cutting of the leaf blade into three parts which also results in enhanced subsequent growth of the marginals. The growth which follows after the release of the marginals from correlative inhibition is accompanied in cut leaf blades with an increased content of endogenous IAA in the period from 12 h to 7 d after leaf isolation when compared with uncut leaf blades. The highest content of endogenous IAA was recorded in the middle section, and the lowest IAA content in the basal section of the leaf blade.     

Branch development in Lupinus angustifolius L.#I. Not all branches have the same potential growth rate

Although the basal and uppermost lateral branches of Lupinus angustifolius L. frequently grow and contribute to yield, buds formed in the axils of leaves 6-12 (referred to as middle buds) rarely grow. This may be due to an inherent limitation of these buds, or some form of apical dominance or competition imposed by the plant. The hypothesis that middle buds have the full capacity to grow, but remain suppressed on intact plants was tested. The main stem apex and buds from the axils of leaves 1 and 8 (bud 1 and bud 8) were excised and cultured on sterile agar. The buds were removed from culture and weighed every 2-3 d for 21 d. The growth rate of apices from the main stem was approximately 5.8 mg d^-1, compared to 2.4 mg d^-1 for bud 1 and 0.9 mg d^-1 for bud 8. Buds in the axils of leaves 6-10 on intact plants were painted six times with a synthetic cytokinin, benzylaminopurine, from 40 d after sowing. This promoted rapid elongation and thickening of these buds, visible as early as 5 d after painting began. The rapid growth of these branches was associated with a reduction in the length of the remaining branches on the plant. However, excision of lower branches did not increase the growth of the middle buds. It is concluded that buds 6-12 of Lupinus angustifolius L. have a partial potential to grow. This potential appears to be limited by innate factors in the bud, and may be structural and/or hormonal. The limitation appears to develop very early in the plant, and potential growth is not modified by subsequent nutrition of the plant.     

THE PHYSIOLOGICAL BASIS OF MORPHOLOGICAL POLARITY IN REGENERATION. I

1. In Bryophyllum calycinum two apical leaves suppress the shoot formation in all the dormant buds situated basally from the leaf; one apical leaf suppresses the shoot formation in the basal buds situated in the same half of the stem where the leaf is, and, if one-half of the petiole of such a leaf is removed, the growth of basal buds in one quadrant of the stem is suppressed. 2. This inhibitory influence of a leaf upon shoot formation in the basal part of a stem is diminished or disappears when the mass of the leaf is reduced below a certain limit. 3. The inhibitory influence of an apical leaf upon the growth of shoots in horizontally suspended stems is greater when the leaf is on the upper than when it is on the lower side of the stem. 4. All these facts suggest the possibility that the inhibitory influence of the leaf upon shoot formation is due to inhibitory substances secreted by the leaf and carried by the sap from the leaf towards the base of the stem. 5. An apical leaf accelerates root formation in the basal part of a stem and this accelerating effect increases with the mass of the leaf. 6. This inhibitory influence of a leaf upon shoot formation and the favoring influence upon root formation in the more basally situated parts of the stem is one of the factors determining the polar character of regeneration.     

THE PHYSIOLOGICAL BASIS OF MORPHOLOGICAL POLARITY IN REGENERATION. II

1. The experiments show that the mass of air roots formed in a stem increases with the mass of the leaf attached to the stem, though it has not been possible to establish an exact mathematical relation between the two masses, owing to unavoidable sources of error. 2. Darkened leaves do not increase the mass of roots formed. 3. In stems suspended horizontally air roots appear on the lower side of the stem, with the exception of the cut end where they usually appear around the whole circumference of the stem. When the lower half of a stem suspended horizontally is cut off, roots are formed on the upper side. It is shown by experiments on leaves suspended horizontally that the more rapidly growing roots and shoots on the lower side inhibit the root and shoot formation in the upper half of such a leaf; and likewise the more rapid formation of roots on the lower side of a horizontally suspended stem seems to account for the inhibition of root formation on the upper side of such a stem. Likewise the more rapid growth of shoots on the upper side of a stem suspended horizontally is likely to inhibit the growth of shoots on the lower side. 4. Each leaf contains in its axil a preformed bud capable of giving rise to a root, which never grows out in the normal stem on account of the inhibitory influence of the normal roots at the base of the plant. These dormant root buds are situated above (apically from) the dormant shoot bud. The apical root buds can be caused to develop into air roots when a piece of stem is cut out from a plant from which the leaves except those in the basal node of the piece are removed. The larger these basal leaves the better the experiments succeed. 5. These apical air roots grow out in a few days, while the roots at the basal end of the stem (which in our experiments dip into water) grow out about a week later. As soon as the basal roots grow out in water they cause the air roots in the more apical region of the stem to dry out and to disappear. 6. In addition to the basal roots, basal nodes have also an inhibitory effect on the growth of the dormant root buds in the apical region of a stem. This is indicated by the fact that a stem with one pair of leaves near the base will form apical air roots more readily when no node is situated on the stem basally from the leaf than if there is a node basally from the leaf.     

The Heterogeneity of Structural and Functional Photosynthetic Characteristics of Mesophyll Chloroplasts in Various Parts of Mature or Senescing Leaf Blade of Two Maize (Zea Mays L.) Genotypes

Differences in ultrastructural parameters of mesophyll cell (MC) chloroplasts, contents of photosynthetic pigments, and photochemical activities of isolated MC chloroplasts were studied in the basal, middle, and apical part of mature or senescing leaf blade of two maize genotypes. A distinct heterogeneity of leaf blade was observed both for structural and functional characteristics of chloroplasts. In both mature and senescing leaves the shape of MC chloroplasts changed from flat one in basal part of leaf to nearly spherical one in leaf apex. The volume density of granal thylakoids decreased from leaf base to apex in both types of leaves examined, while the amount of intergranal thylakoids increased in mature leaves but decreased in senescing leaves. The most striking heterogeneity was found for the quantity of plastoglobuli, which strongly increased with the increasing distance from leaf base. The differences in chloroplast ultrastructure were accompanied by differences in other photosynthetic characteristics. The Hill reaction activity and activity of photosystem 1 of isolated MC chloroplasts decreased from leaf base to apex in mature leaves. Apical part of senescing leaf blade was characterised by low contents of chlorophyll (Chl) a and Chl b, whereas in mature leaves, the content of Chls as well as the content of total carotenoids (Car) slightly increased from basal to apical leaf part. This was reflected also in the ratio Chl (a+b)/total Car; the ratio of Chl a/b did not significantly differ between individual parts of leaf blade. Both genotypes examined differed in the character of developmental gradient observed along whole length of leaf blade.     

Bud formation in leaves,leaf fragments and midrib pieces of Heloniopsis orientalis (Liliaceae)

Summary Isolated leaves, leaf fragments and pieces of the midrib portion devoid of lamina, of Heloniopsis orientalis were grown on an inorganic nutrient medium without organic nutrients and growth regulators in order to investigate their regenerative ability. Bud formation in intact, attached leaves occurs only at the tip, in isolated leaves at the tip and the base, whereas leaf fragments cut transversely at a distance from the tip and isolated midrib pieces form numerous shoot buds in a random distribution. Lamina fragments lacking midrib frequently fail to regenerate even after a long time of culture. It is suggested that endogeneous growth regulators in the leaf, especially the vascular tissues, play an important role in bud initiation. Very young leaves of Heloniopsis are capable forming buds and roots when isolated from the mother plants.     

Relative importance of nodal roots and apical buds in the control of branching in Trifolium repens L.

Clonal species are characterised by having a growth form in which roots and shoots originate from the same meristem so that adventitious nodal roots form close to the terminal apical bud of stems. The nature of the relationship between nodal roots and axillary bud growth was investigated in three manipulative experiments on cuttings of a single genotype of Trifolium repens. In the absence of locally positioned nodal roots axillary bud development within the apical bud proceeded normally until it slowed once the subtending leaf had matured to be the second expanded leaf on the stem. Excision of apical tissues indicated that while there was no apical dominance apparent within fully rooted stems and very little in stems with 15 or more unrooted nodes, the outgrowth of the two most distal axillary buds was stimulated by decapitation in stems with intermediate numbers of unrooted nodes. Excision of the basal branches from stems growing without local nodal roots markedly increased the length and/or number of leaves on 14 distally positioned branches. The presence of basal branches therefore prevented the translocation of root-supplied resources (nutrients, water, phytohormones) to the more distally located nodes and this caused the retardation in the outgrowth of their axillary buds. Based on all three experiments we conclude that the primary control of bud outgrowth is exerted by roots via the acropetal transport of root-supplied resources necessary for axillary bud outgrowth and that apical dominance plays a very minor role in the regulation of axillary bud outgrowth in T. repens.     

Another evidence for inhibitory effect of auxin in adventitious bud formation of decapitated flax (Linum usitatissimum L.) seedlings

Adventitious buds were formed on the hypocotyls of decapitated flax seedlings. Scanning electron and light microscopic examinations of hypocotyls showed that epidermal cells divided to produce meristematic spots from which several leaf primordia were formed. Between leaf primordia and the original vascular tissues of hypocotyls, new xylem cells were formed which connected them. About 10, 30 and 60% of adventitious buds were formed on upper, middle and basal parts of hypocotyls of decapitated seedlings, respectively. Removal of apical meristem together with longer hypocotyl zero to four cm long below the apical meristem) induced higher percentage of adventitious bud formation in the remaining hypocotyl. When the entire hypocotyl was cut into 16 segments (0.25 cm each) and these segments were cultured on MS medium containing 3% sucrose and 0.8% agar, adventitious buds were mainly formed in the lowest five segments. These results suggested that there was a gradient of inhibitory factor(s) from apical to basal part of hypocotyl with respect to adventitious bud formation. Auxin transport inhibitors, morphactin and TIBA induced adventitious bud formation on intact seedlings by suppressing the basipetal movement of auxin.     

Endogenous levels of abscisic acid,indole-3-acetic acid and benzyladenine during in vitro bud growth induction of Wild cherry (Prunus avium L.)

Levels of endogenous growth substances (abscisic acid: ABA; indole-3-acetic acid: IAA) and applied benzyladenine (BA) were quantified during the eight first days of in vitro propagation of Wild Cherry (Prunus avium L.). Axillary buds from the middle part of the explants started to grow at day 2, thus were released from apical dominance. Hormone levels were measured in the apical, median and basal parts of the explants using an avidin-biotin based enzyme-linked immunosorbent assay (ELISA) after a purification of the extracts by high performance liquid chromatography (HPLC). All hormones showed rapid and considerable changes during the first eight days of growth. Exogenous IBA was probably transformed into IAA mainly in the basal part of the explant, and BA penetrated quickly. ABA levels were transiently enhanced in the apical part of the explants bearing young leaves. These phenomena are discussed in connection with the axillary bud reactivation.     

Bud Structure in Relation to Shoot Morphology and Position on the Vegetative Annual Shoots of Juglans regia L. (Juglandaceae)

The length and basal diameter of all lateral and terminal budsof vegetative annual shoots of 7-year-oldJuglans regia treeswere measured. All buds were dissected and numbers of cataphylls,embryonic leaves and leaf primordia were recorded. Each axillarybud was ranked according to the position of its associated leaffrom the apex to the base of its parent shoot. Bud size andcontent were analysed in relation to bud position and were comparedwith the size and number of leaves of shoots in equivalent positionswhich extended during the following growing season. Length andbasal diameter of axillary buds varied according to their positionon the parent shoot. Terminal buds contained more embryonicleaves than any axillary bud. The number of leaves was smallerfor apical and basal axillary buds than for buds in intermediatepositions on the parent shoot only. All new extended shootswere entirely preformed in the buds that gave rise to them.Lateral shoots were formed in the median part of the parentshoot. These lateral shoots derived from buds which were largerthan both apical and basal ones. Copyright 2001 Annals of BotanyCompany Juglans regia L., Persian walnut tree, branching pattern, preformation, bud content, shoot morphology     

Diurnal oscillatory movements of growing leaves of tobacco

The analysis of diurnal oscillatory movements of tobacco leaves was used in the diagnosis of viral infection of plants. The oscillatory helices circumscribed by a growing leaf of a healthy plant were regular, but some deviations, particularly in the transition points, were recorded. In order to make clear the cause of these irregularities of trajectory, the course of elongation of leaf petiole and blade in relation to localization and shift of zones of elongation during ontogenesis was analysed. The present analysis is similar to that described by the author's earlier experiments with pea roots. Oscillatory curves circumscribed by petiole, projected on a horizontal plane, were compared with curves circumscribed by the blade tip. The analysis of the leaves of different ages enabled us to study this process in dependence on growth rate. It was confirmed that oscillations are a result of elongation; the extent of oscillations is quantitatively dependent on the growth rate. An analysis of the zones of growth showed that in petiole the active meristems are localized near to its base while in the leaf lamina they move gradually during the ontogenesis from the apical to the basal part of the leaf blade. Active meristems of young rapidly growing leaves are localized approximately in the middle of the blade while those of old leaves were found in close proximity to the base of the lamina. The growth rate of petiole can be expressed in hundreds of mm per hour (4.8×10^?2 mm h^?1); half of this value was recorded for its apical part. The growth rate of leaf blade was found approximately ten times higher (3.2×10^?1 mm h^?1). The oscillatory movements of growing leaf consists of two integrate components: of oscillations originating in the base of the petiole and of oscillations of leaf blade the centrum of which is localized in the basal third of the blade. The arrangement of the experiments did not enable us to determine to what extent the phototropic response of leaf blade participates in leaf movements. The movements of leaves of an intact plant are evidently affected by rhythmic stem oscillations. Stem is an integral part of a system which participates in the transfer of information in plants.     

DEVELOPMENT AND PHYLLOTAXIS OF THE VEGETATIVE AXILLARY BUD OF MICHELIA FUSCATA

Tucker, Shirley C. (Northwestern U., Evanston, III.) Development and phyllotaxis of the vegetative axillary bud of Michelia fuscata . Amer. Jour. Bot. 50(7): 661–668. Illus. 1963.—The vegetative axillary buds of Michelia fuscala are dorsiventrally symmetrical with 2 ranks of alternately produced leaves. The direction of the ontogenetic spiral in each of these buds is related both to the symmetry of the supporting branch and to the position of the bud along the branch. On a radially symmetrical branch, all the axillary buds are alike—all clockwise, for example. But in a dorsiventrally organized branch the symmetry alternates from clockwise in 1 axillary bud to counterclockwise in the next bud along the axis. Leaf initiation and ontogeny of the axillary apical meristem conform with those of the terminal vegetative bud. The axillary bud arises as a shell zone in the second leaf axil from the terminal meristem. During this process the axillary apex develops a zonate appearance. The acropetally developing procambial supply of the axillary bud consists wholly of leaf traces. At the nodal level the bud traces diverge from the same gap as the median bundle trace of the subtending leaf. Only the basal 1–2 axillary buds which form immediately after the flowers elongate each year, while the majority remains dormant with 3 leaves or fewer.     

Shoot elongation,leaf demography and bud formation in relation to branch position on Larix laricina saplings

Summary Shoot development was investigated on branches of Larix laricina (Du Roi) K. Koch trees growing in their 8th year in two plantations and in a natural stand approximately 12 years old. Expansion of throughout-crown series of short and long shoots was measured weekly, and later colour change and natural fall of leaves were assessed. Similar shoots were collected at intervals and dissected, the long shoots by 25-leaf segments. Leaf area and weight, as well as time of bud formation, were determined. Increasing acropetal trends were evident in time to bud burst: duration of short-shoot leaf-cluster expansion; size of leaf clusters and number, area and weight of leaves per cluster; duration and rate of long-shoot elongation; number, area and weight of leaves on long shoots; time to terminal-bud formation on long shoots. Along each long shoot, stem and leaf elongation and lateral-axis formation progressed acropetally. Lateral axes were most numerous on second to fourth 25-leaf segments. On longer shoots, some axes in middle segments developed as sylleptic short shoots rather than as lateral buds. Leaves of short shoots and basal leaves on long shoots turned yellow and abscissed sooner than axial leaves on long shoots. Colour change and loss among axial leaves were acropetal along shoots and up the crown. Thus, last-formed leaves, in axils of some of which lastformed lateral buds occurred, were held longest.     

Effects of Wounding on Adventitious Bud Formation in Torenia Stem Segments Cultured In Vitro

In in vitro cultured stem segments of Torenia fournieri Lind.,the formation of adventitious buds can be induced when the culturemedium contains cytokinin. When long stem segments (2.0 cm ormore) were cultured with cytokinin, a large number of buds wereformed in the marginal regions, namely, within the limits of0.5 cm from the cut ends of explants, while only a few budswere initiated in the middle part of the explants. If a slightinjury was made transversely with a scalpel in the central partof an explant, a significant increase in the number of budswas noted within the limits of 0.5 cm from the wound site. Whena wounding treatment was given lengthwise to an explant, a largenumber of adventitious buds were formed over the entire surfaceof the explant compared to the control. Excision itself of explantsfrom mother plants and the additional wounding given to theexplants seemed to trigger the induction of adventitious buddifferentiation in Torenia stem segments. These wounding treatmentsdid not affect the uptake into explants and/or the distributionpattern of radioactive benzyladenine applied to the culturemedium. Key words: Torenia fournieri, Adventitious bud formation, Cytokinin, Wounding     

INDETERMINATE GROWTH OF LEAVES IN GUAREA (MELIACEAE): A TWIG ANALOGUE

Leaves of seed plants are generally characterized as organs of determinate growth. In this regard, Guarea and related genera seem unusual in that the pinnately compound leaves of these plants contain a bud at their tip from which new pinnae expand from time to time. Previous studies (based upon superficial examinations of leaf-tip buds) have produced contradictory conclusions regarding how long the leaf apex remains meristematic and produces new pinna primordia. In order to determine whether leaf development in Guarea is truly indeterminate, we microscopically examined leaf-tip buds of G. guidonia and G. glabra. In both species, the leaf apex remains meristematic and continues to produce new pinna primordia as the leaf ages. Unexpanded leaves of G. guidonia contained an average of 23 pinna primordia, while the oldest leaves we examined had initiated an average of 44 total pinnae. In G. glabra, unexpanded leaves contained 8 pinnae, whereas an average of 28 pinnae had been initiated on the oldest leaves. These results indicate that leaf development in Guarea is truly indeterminate. Periodic examination of individual intact leaves indicated that the leaves commonly continue their growth for 2 or more years (observed maximum = 51 months). As new leaflets are initiated at the shoot apex (and subsequently expand in rhythmic flushes), older (basal) leaflets may abscise. In addition, the petiole and rachis of the leaf thicken and become woody as a result of the activity of a vascular cambium. Guarea leaves therefore seem to function as the analogue of a typical twig (stem) in general habit as well as in their indeterminate apical growth and secondary thickening.