The Morphogenesis of Apple Buds: II. The Development of the Bud

The formation of the terminal resting bud of apple spurs wasstudied on untreated and defoliated spurs by dissection of budscollected at intervals during the season. A distinctive patternof bud development was found in all treatments, the principalfeatures being firstly the steady rate over long periods ofthe season at which successive primordia began or completedbud-scale development, and secondly a change in rate of initiationof bud-scales several weeks after the resting bud had begunto form. Bud-scale initiation was invariably delayed in primordia inthe early period of bud development, when primordia adjacentto the apical meristem were affected directly by the foliage.The rate at which the number of mature bud-scales in the budincreased during the season was not related to the extent offoliar development, but appeared to depend upon the degree towhich the bud, during its early development, was affected bythe foliage. Whilst defoliation of spurs showed that the formation of bud-scaleswas dependent upon an effect of the foliage, no simple relationshipwas found. The number and size of leaves on defoliated spurswas much less than on untreated spurs, but the pattern of buddevelopment was very similar. Since the morphogenetic effectof foliage is known to vary with its age, the constant rateof formation of bud-scales during the season was consideredto be evidence of a second factor in the bud which counteractedvariations in effect of the foliage. A change in rate of increasein number of bud-scales could only occur therefore if the effectivelevel of one of these factors was altered independently of theother. This, it is suggested, is brought about by the accumulationof a further factor in the bud-scales, at a rate determinedduring the early formation of each primordium. From the similaritybetween the effects of these hypothetical factors and thoseof growth-regulating substances extracted from or applied tobuds of fruit trees it is thought that the second factor maybe a gibberellin-like substance, and the bud-scale factor agrowth inhibitor.     

Apple Fruit Bud Development and Growth; Analysis and an Empirical Model

Analysis of the information available on apple bud developmentand growth after dormancy leads to an empirical model of growthto full bloom. The analysis and model are set in the frameworkof the physiological mechanisms considered to be responsiblefor dormancy and subsequent bud growth. It is necessary to introducean arbitrary ‘growth unit’ scale to describe theseprocesses quantitatively, which is done by the equation G = A/(I+be^–k(I).P) where G and A are in growth units, the value of k is controlledby a dormancy index I and P is a temperature summation. Themodel fulfils the main requirements laid down for it and thevalues of P at full bloom, derived from controlled environmentwork and field observations, are very similar.     

苹果花芽孕育机理的探讨

花芽孕育期,以环剥、多效唑＋环剥和ＧＡ３处理“红富士”苹果树,成花效应以多效唑＋环剥处理为最高（＋６８４％）。花芽孕育起动时,促花处理的短枝顶芽内ＩＡＡ、ＺＲ含量和ＩＡＡ／ＧＡｓ、ＺＲ／ＧＡｓ的比值均明显提高。在花芽孕育完成过程中,促花处理的短枝叶内淀粉含量、Ｃ／Ｎ比,芽内ＺＲ、ＩＡＡ含量和ＺＲ／ＧＡｓ明显提高,ＧＡｓ则明显下降。     

Mechanisms of Side Branching and Tip Splitting in a Model of Branching Morphogenesis

Recent experimental work in lung morphogenesis has described an elegant pattern of branching phenomena. Two primary forms of branching have been identified: side branching and tip splitting. In our previous study of lung branching morphogenesis, we used a 4 variable partial differential equation (PDE), due to Meinhardt, as our mathematical model to describe the reaction and diffusion of morphogens creating those branched patterns. By altering key parameters in the model, we were able to reproduce all the branching styles and the switch between branching modes. Here, we attempt to explain the branching phenomena described above, as growing out of two fundamental instabilities, one in the longitudinal (growth) direction and the other in the transverse direction. We begin by decoupling the original branching process into two semi-independent sub-processes, 1) a classic activator/inhibitor system along the growing stalk, and 2) the spatial growth of the stalk. We then reduced the full branching model into an activator/inhibitor model that embeds growth of the stalk as a controllable parameter, to explore the mechanisms that determine different branching patterns. We found that, in this model, 1) side branching results from a pattern-formation instability of the activator/inhibitor subsystem in the longitudinal direction. This instability is far from equilibrium, requiring a large inhomogeneity in the initial conditions. It successively creates periodic activator peaks along the growing stalk, each of which later on migrates out and forms a side branch; 2) tip splitting is due to a Turing-style instability along the transversal direction, that creates the spatial splitting of the activator peak into 2 simultaneously-formed peaks at the growing tip, the occurrence of which requires the widening of the growing stalk. Tip splitting is abolished when transversal stalk widening is prevented; 3) when both instabilities are satisfied, tip bifurcation occurs together with side branching.     

The Morphogenesis of Apple Buds: IV. The Effect of Fruit

The course of development of spur buds on fruiting trees of‘biennial’ and ‘regular’ apple varietieswas followed throughout the season by dissection of successivesamples of buds and the effect on bud development of de-fruitingtrees at two different times in the season was recorded. In the biennially fruiting variety Miller's Seedling, the patternof bud development on fruiting trees was very similar to thatin buds on non-fruiting trees which failed to form flowers followingdefoliation, as were the number and sizes of leaves precedingthe bud in each instance. The failure to form flowers was associatedwith the occurrence of an 18-day plastochrone in buds, and itwas concluded that on fruiting trees it was not due to a competitiveeffect of the fruit for nutrients. This long plastochrone was not found in buds of de-blossomedtrees of Miller's Seedling, and in trees de-fruited later inthe season the buds immediately broke into a new flush of growthand at the same time the plastochrone was shortened. These resultssuggest that the long plastochrone was due to the inhibitoryeffect of fruit on the older primordia of the bud, an effectwhich did not occur until after the resting buds had begun toform. A phase with an 18-day plastochrone was also found inbuds of another biennial variety, Laxton's Superb, but not inthose of the regularly fruiting variety Sunset. Developing fruitlets of biennial varieties caused bud-scalesto form sooner and hastened their rate of development, possiblydue to changes induced in the levels of a gibberellin-like apexfactor in the buds. The rate of increase in number of bud-scalesin the bud appeared to depend upon the extent to which the budwas affected by the primary leaves of the flower cluster andthose of other clusters.     

The Morphogenesis of Apple Buds: III. The Inception of Flowers

The early stages in the change from vegetative to reproductivedevelopment of apple spur terminal buds were followed by dissectionof buds from untreated trees, and from trees defoliated at differenttimes in the season. A change in the development of the leafprimordia occurred when there were approximately eight in thebud. This was followed by the development of bracts, which appearedto be necessary for the formation of actual flower parts. Leafprimordia tend to inhibit this process. Whereas their effectupon the apical meristem was subsequently reduced by the formationof bracts, so that eventually a terminal flower formed, theireffect upon the lower lateral meristems was unaltered. Thesemeristems therefore remained in a vegetative state. In addition to the number of leaf primordia in the bud, thedegree of dormancy may be an important factor in determiningthe onset of flowering. Since the number of leaf primordia invegetative buds at the end of the season is eight, the spatialdistribution of primordia on the main axis of the bud and theirvascular connexions might have a decisive effect on bud development.This was related to the effect of older primordia in the budupon the development of younger ones. In buds in which theseolder primordia were inhibited by foliage, etc., i.e. thosewith a long plastochrone, no effects were observed upon thedevelopment of younger primordia and the buds remained vegetative. Whilst correlative inhibition of buds thus affected their abilityto form flowers, there is no evidence of a critical leaf areafor flowering. Flowering in apple buds is more likely to bedue to the removal of factors inhibiting reproductive developmentthan to the synthesis of a specific flower inducing substanceas such.     

Growth Based Morphogenesis of Vertebrate Limb Bud

Many genes and their regulatory relationships are involved in developmental phenomena. However, by chemical information alone, we cannot fully understand changing organ morphologies through tissue growth because deformation and growth of the organ are essentially mechanical processes. Here, we develop a mathematical model to describe the change of organ morphologies through cell proliferation. Our basic idea is that the proper specification of localized volume source (e.g., cell proliferation) is able to guide organ morphogenesis, and that the specification is given by chemical gradients. We call this idea “growth-based morphogenesis.” We find that this morphogenetic mechanism works if the tissue is elastic for small deformation and plastic for large deformation. To illustrate our concept, we study the development of vertebrate limb buds, in which a limb bud protrudes from a flat lateral plate and extends distally in a self-organized manner. We show how the proportion of limb bud shape depends on different parameters and also show the conditions needed for normal morphogenesis, which can explain abnormal morphology of some mutants. We believe that the ideas shown in the present paper are useful for the morphogenesis of other organs.     

The Morphogenesis of Apple Buds: I. The Activity of the Apical Meristem

The mechanism controlling the production of primordia by theapical meristem of apple buds has been studied by means of defoliationexperiments and the dissection of buds. It is shown that theapical meristem of the bud passes through a series of comparativelystable phases of activity, changing relatively abruptly fromone phase to another. The relation of the activity of the apicalmeristem to the development of the bud and foliage is discussed,and it is concluded that the rate of production of primordiais controlled by the younger leaf primordia in the bud, whichmay themselves be controlled by the foliage.     

The Anatomy of the Developing Bud Union and its Relationship to Dwarfing in Apple

Light microscopy has been used to study the effect of dwarfingand semi-dwarfing apple rootstocks on the early developmentof bud-unions with 'Gala', and the anatomy of 2-year-old bud-unionsbetween 'Bramley' and the same rootstocks. The bridging of thecut edges of the cambia of bud and rootstock was achieved bydifferentiation of callus formed at an early stage in budding.New cambial cells were aligned at right angles to the pre-existingcambia, with their long axes horizontal. Subsequently-formedxylem adopted this arrangement, so that fibres and vessels werearranged obliquely to the axis of the stem. At the interfacebetween the bud and dwarfing rootstocks vessels with smallerthan normal diameter were formed, indicating the presence ofelevated levels of auxin in this region. In addition, littlexylem was produced in the adjacent rootstock tissue. In thecase of semi-dwarfing rootstocks, the rootstock produced normalxylem after a brief interruption. We suggest that failure ofauxin to cross the bud-union interface in the case of the dwarfingrootstocks leads to reduced rootstock xylem formation, and hencea poor supply of water and minerals to the scion, and this underliesthe dwarfing effect.Copyright 1994, 1999 Academic Press Apple, budding, dwarfing, anatomy, graft union     

Multi-Scale Modeling in Morphogenesis: A Critical Analysis of the Cellular Potts Model

Cellular Potts models (CPMs) are used as a modeling framework to elucidate mechanisms of biological development. They allow a spatial resolution below the cellular scale and are applied particularly when problems are studied where multiple spatial and temporal scales are involved. Despite the increasing usage of CPMs in theoretical biology, this model class has received little attention from mathematical theory. To narrow this gap, the CPMs are subjected to a theoretical study here. It is asked to which extent the updating rules establish an appropriate dynamical model of intercellular interactions and what the principal behavior at different time scales characterizes. It is shown that the longtime behavior of a CPM is degenerate in the sense that the cells consecutively die out, independent of the specific interdependence structure that characterizes the model. While CPMs are naturally defined on finite, spatially bounded lattices, possible extensions to spatially unbounded systems are explored to assess to which extent spatio-temporal limit procedures can be applied to describe the emergent behavior at the tissue scale. To elucidate the mechanistic structure of CPMs, the model class is integrated into a general multiscale framework. It is shown that the central role of the surface fluctuations, which subsume several cellular and intercellular factors, entails substantial limitations for a CPM''s exploitation both as a mechanistic and as a phenomenological model.     

Effect of Purine Analogs and Their Interactions with Benzyladenine on Bud Burst and Shoot Growth in Apple Bud Explants

The purine analogs 8-azaadenine, 8-azaguanine, 4-aminopyrazolo [3,4-d] pyrimidine, 2-fluoroadenine and 6-methylpurine inhibited bud burst in non-dormant vegetative axillary bud explants obtained from apple (Malus domestica Borkh.) seedlings. The reversal of inhibition was attempted by incorporating increasing concentrations of benzyladenine (BA) in the culture medium. The inhibition caused by 8-azaadenine and 8-azaguanine was reduced especially when BA was added to the media at equimolar ratios along with the inhibitors. The inhibition due to 4-aminopyrazolo [3,4-d] pyridine was reduced only with equimolar BA, BA was ineffective or only minimally effective in reducing inhibition due to 2-fluoroadenine and 6-methypurine. Short pretreatment with 8-azaadenine and 8-azaguinine failed to cause any inhibition when explants were subsequently transferred to BA -containing medium. The effectiveness of 2-fluoroadenine and 6-methylpurine was maintained or only slightly reduced, indicating their long-term inhibitory action.     

Whole-mount Immunohistochemical Analysis for Embryonic Limb Skin Vasculature: a Model System to Study Vascular Branching Morphogenesis in Embryo

Whole-mount immunohistochemical analysis for imaging the entire vasculature is pivotal for understanding the cellular mechanisms of branching morphogenesis. We have developed the limb skin vasculature model to study vascular development in which a pre-existing primitive capillary plexus is reorganized into a hierarchically branched vascular network. Whole-mount confocal microscopy with multiple labelling allows for robust imaging of intact blood vessels as well as their cellular components including endothelial cells, pericytes and smooth muscle cells, using specific fluorescent markers. Advances in this limb skin vasculature model with genetic studies have improved understanding molecular mechanisms of vascular development and patterning. The limb skin vasculature model has been used to study how peripheral nerves provide a spatial template for the differentiation and patterning of arteries. This video article describes a simple and robust protocol to stain intact blood vessels with vascular specific antibodies and fluorescent secondary antibodies, which is applicable for vascularized embryonic organs where we are able to follow the process of vascular development.Download video file.^{(59M, mov)}     

Pattern Formation in a Nonlinear Membrane Model for Epithelial Morphogenesis

A theoretical model is presented for pattern formation in an epithelium. The epithelial model consists of a thin, incompressible, viscoelastic membrane on an elastic foundation (substrate), with the component cells assumed to have active contractile properties similar to those of smooth muscle. The analysis includes the effects of large strains and material nonlinearity, and the governing equations were solved using finite differences. Deformation patterns form when the cells activate while lying on the descending limb of their total (active + passive) stress-stretch curve. Various one-dimensional and two-dimensional simulations illustrate the effects of spatial and temporal variations in passive stiffness, as well as the effects of foundation stiffness and stretch activation. The model can be used to examine the mechanical aspects of pattern formation in morphogenetic processes such as angiogenesis and myocardial trabeculation.     

Differentiation of Apical Bud Cells in a Newly Developed Apical Bud Transplantation Model Using GFP Transgenic Mice as Donor

Rodent mandibular incisors have a unique anatomical structure that allows teeth to grow throughout the lifetime of the rodent. This report presents a novel transplantation technique for studying the apical bud differentiation of rodent mandibular incisors. Incisal apical end tissue with green fluorescent protein from transgenic mouse was transplanted to wild type mice, and the development of the transplanted cells were immunohistologically observed for 12 weeks after the transplantation. Results indicate that the green fluorescent apical end tissue replaced the original tissue, and cells from the apical bud differentiated and extended toward the incisal edge direction. The immunostaining with podoplanin also showed that the characteristics of the green fluorescent tissue were identical to those of the original. The green fluorescent cells were only found in the labial side of the incisor up to 4 weeks. After 12 weeks, however, they were also found in the lingual side. Here the green fluorescent cementocyte-like cells were only present in the cementum close to the dentin surface. This study suggests that some of the cells that form the cellular cementum come from the apical tissue including the apical bud in rodent incisors.     

Acetabularia: A Unicellular Model for Understanding Subcellular Localization and Morphogenesis during Development

Acetabularia acetabulum is the organism that provided the first compelling experimental evidence both for the role of an organelle whose function was unknown, the nucleus, and for the existence of "morphogenetic substances," the behavior of which presaged the discovery of mRNA in other organisms. This giant unicell holds special appeal as a model system, because the contribution of its diploid nucleus to cellular processes can be assessed using simple amputation and grafting experiments and because it lends itself to a wide range of methods in cell, molecular, and developmental biology. It remains an excellent model system for understanding how body regions are functionally and structurally distinguished from each other without cellular compartmentation. Advances in genetics (that is, mutant selection and analysis, large-scale transformation) will greatly increase the power of this system to address fundamental questions in development and morphogenesis. We discuss strengths and weaknesses of the system and outline the body of knowledge that would make the system more powerful and broadly appealing.