Developmental Anatomy of Axillary Meristems of Araucaria cunninghamii Released from Apical Dominance Following Shoot Apex Decapitation In vitro and In vivo

1989 ◽  
Vol 150 (4) ◽  
pp. 369-377 ◽  
Author(s):  
G. E. Burrows
1988 ◽  
Vol 36 (6) ◽  
pp. 665 ◽  
Author(s):  
GE Burrows ◽  
DD Doley ◽  
RJ Haines ◽  
DG Nikles

Stem segments with 3-5 leaf axils, excised from the upper portion of the mainstem of 2-year-old hoop pine (Araucaria cunninghamii Aiton ex D. Don) seedlings, produced orthotropic buds from the concealed axillary meristems when cultured on a basal medium (BM) of half-strength Murashige and Skoog (MS) inorganic salts, the medium level of growth factors and amino acids of de Fossard, 20 g L sucrose and 6.5 g/L agar. This procedure was also successful with A. balansae, A. bidwillii, A. colurnnaris, A. hunsteinri, A. luxurians, A. montana, A. rulei, A. scopulorum and Agathis robusta and with stem segments from orthotropic coppice shoots of juvenile morphology collected from the stumps of 20-year old hoop pines felled near ground level. The hoop pine explants were highly sensitive to cytokinin; 1 μM and 10 μM 6-benzylaminopurine caused the formation of distorted buds and total inhibition of bud development respectively. Lofier concentrations (0.001-0.1 μM ) did not noticeably influence bud formation or development. A low rate of multiplication was induced by reculturing the stem segments after the excision of the initial shoots. New buds developed in the leaf axils of that part of the initial shoot which remained attached to the primary stem explant. Shoots derived from seedling and coppice cultures of hoop pine and seedling cultures of Agathis robusta rooted in vitro on BM + 0.1-10.0 μM indole-3-butyric acid (IBA), but with only 5-20% success. Up to 80% rooting was obtained if both hoop pine shoot types (i. e. from seedling and coppice cultures) were cultured on modified BM (quarter strength MS salts, 10 μM IBA plus no agar) for 2 weeks, before being transferred to a mixture of non-sterile peat and perlite or vermiculite and perlite, maintained under a high humidity (90-95%). Plantlets were subsequently transferred to normal glasshouse conditions and then to the field with less than 5% mortality. Thus hoop pine can be added to the relatively small number of conifers for which the capacity to micropropagate juvenile and mature plants and successfully establish their clones in the field has been demonstrated.


2011 ◽  
Vol 72 (4) ◽  
pp. 295-302 ◽  
Author(s):  
Tomasz Pniewski ◽  
Joanna Wachowiak ◽  
Józef Kapusta ◽  
Andrzej B. Legocki

The complete protocol for regeneration and long-term micropropagation of several Polish cultivars of pea (<em>Pisum sativum </em>L.) has been elaborated. The shoots were the most likely regenerated via de novo organogenesis. The adventitious buds formed in callus derived from cotyledons tissue adjacent to the axillary meristems of immature embryos. All cultivars' calli regenerated several shoots per explant on the MS medium supplemented with B5 vitamins and 4.5 mgl<sup>-1</sup> of BAP, however some differences in regeneration capacity among cultivars were observed. The plantlets were subsequently micropropagated with slightly higher efficiency and preserving a good viability over the long-term culture on a medium containing 2.0 mgl<sup>-1</sup> than one with 4.5 mgl<sup>-1</sup> of BAP. The additional step of the pre-conditioning culture of multiplicated shoots on a medium with very low BAP concentration i.e. 0.02 mgl<sup>-1</sup> was applied and appeared to be beneficial before rooting in vitro or grafting. The modified MS-derived medium with the half-strength of MS macroelements but with the full original dose of calcium and supplemented with B5 vitamins and 1.0 mgl<sup>-1</sup> of NAA was developed for effective rooting. The shoots were also sufficiently transferred into ex vitro conditions using grafting. The majority of the regenerated plants had adapted to in vivo conditions in a greenhouse and subsequently has set seeds. The presented protocol provides relatively efficient rate of de novo pea regeneration and would be useful for <em>Agrobacterium</em>-mediated transformation purposes.


Author(s):  
E. J. Kollar

The differentiation and maintenance of many specialized epithelial structures are dependent on the underlying connective tissue stroma and on an intact basal lamina. These requirements are especially stringent in the development and maintenance of the skin and oral mucosa. The keratinization patterns of thin or thick cornified layers as well as the appearance of specialized functional derivatives such as hair and teeth can be correlated with the specific source of stroma which supports these differentiated expressions.


Author(s):  
M.J. Murphy ◽  
R.R. Price ◽  
J.C. Sloman

The in vitro human tumor cloning assay originally described by Salmon and Hamburger has been applied recently to the investigation of differential anti-tumor drug sensitivities over a broad range of human neoplasms. A major problem in the acceptance of this technique has been the question of the relationship between the cultured cells and the original patient tumor, i.e., whether the colonies that develop derive from the neoplasm or from some other cell type within the initial cell population. A study of the ultrastructural morphology of the cultured cells vs. patient tumor has therefore been undertaken to resolve this question. Direct correlation was assured by division of a common tumor mass at surgical resection, one biopsy being fixed for TEM studies, the second being rapidly transported to the laboratory for culture.


Author(s):  
Raul I. Garcia ◽  
Evelyn A. Flynn ◽  
George Szabo

Skin pigmentation in mammals involves the interaction of epidermal melanocytes and keratinocytes in the structural and functional unit known as the Epidermal Melanin Unit. Melanocytes(M) synthesize melanin within specialized membrane-bound organelles, the melanosome or pigment granule. These are subsequently transferred by way of M dendrites to keratinocytes(K) by a mechanism still to be clearly defined. Three different, though not necessarily mutually exclusive, mechanisms of melanosome transfer have been proposed: cytophagocytosis by K of M dendrite tips containing melanosomes, direct injection of melanosomes into the K cytoplasm through a cell-to-cell pore or communicating channel formed by localized fusion of M and K cell membranes, release of melanosomes into the extracellular space(ECS) by exocytosis followed by K uptake using conventional phagocytosis. Variability in methods of transfer has been noted both in vivo and in vitro and there is evidence in support of each transfer mechanism. We Have previously studied M-K interactions in vitro using time-lapse cinemicrography and in vivo at the ultrastructural level using lanthanum tracer and freeze-fracture.


Author(s):  
D. Reis ◽  
B. Vian ◽  
J. C. Roland

Wall morphogenesis in higher plants is a problem still open to controversy. Until now the possibility of a transmembrane control and the involvement of microtubules were mostly envisaged. Self-assembly processes have been observed in the case of walls of Chlamydomonas and bacteria. Spontaneous gelling interactions between xanthan and galactomannan from Ceratonia have been analyzed very recently. The present work provides indications that some processes of spontaneous aggregation could occur in higher plants during the formation and expansion of cell wall.Observations were performed on hypocotyl of mung bean (Phaseolus aureus) for which growth characteristics and wall composition have been previously defined.In situ, the walls of actively growing cells (primary walls) show an ordered three-dimensional organization (fig. 1). The wall is typically polylamellate with multifibrillar layers alternately transverse and longitudinal. Between these layers intermediate strata exist in which the orientation of microfibrils progressively rotates. Thus a progressive change in the morphogenetic activity occurs.


Author(s):  
Conly L. Rieder ◽  
S. Bowser ◽  
R. Nowogrodzki ◽  
K. Ross ◽  
G. Sluder

Eggs have long been a favorite material for studying the mechanism of karyokinesis in-vivo and in-vitro. They can be obtained in great numbers and, when fertilized, divide synchronously over many cell cycles. However, they are not considered to be a practical system for ultrastructural studies on the mitotic apparatus (MA) for several reasons, the most obvious of which is that sectioning them is a formidable task: over 1000 ultra-thin sections need to be cut from a single 80-100 μm diameter egg and of these sections only a small percentage will contain the area or structure of interest. Thus it is difficult and time consuming to obtain reliable ultrastructural data concerning the MA of eggs; and when it is obtained it is necessarily based on a small sample size.We have recently developed a procedure which will facilitate many studies concerned with the ultrastructure of the MA in eggs. It is based on the availability of biological HVEM's and on the observation that 0.25 μm thick serial sections can be screened at high resolution for content (after mounting on slot grids and staining with uranyl and lead) by phase contrast light microscopy (LM; Figs 1-2).


Author(s):  
Robert J. Carroll ◽  
Marvin P. Thompson ◽  
Harold M. Farrell

Milk is an unusually stable colloidal system; the stability of this system is due primarily to the formation of micelles by the major milk proteins, the caseins. Numerous models for the structure of casein micelles have been proposed; these models have been formulated on the basis of in vitro studies. Synthetic casein micelles (i.e., those formed by mixing the purified αsl- and k-caseins with Ca2+ in appropriate ratios) are dissimilar to those from freshly-drawn milks in (i) size distribution, (ii) ratio of Ca/P, and (iii) solvation (g. water/g. protein). Evidently, in vivo organization of the caseins into the micellar form occurs in-a manner which is not identical to the in vitro mode of formation.


Author(s):  
U. Aebi ◽  
L.E. Buhle ◽  
W.E. Fowler

Many important supramolecular structures such as filaments, microtubules, virus capsids and certain membrane proteins and bacterial cell walls exist as ordered polymers or two-dimensional crystalline arrays in vivo. In several instances it has been possible to induce soluble proteins to form ordered polymers or two-dimensional crystalline arrays in vitro. In both cases a combination of electron microscopy of negatively stained specimens with analog or digital image processing techniques has proven extremely useful for elucidating the molecular and supramolecular organization of the constituent proteins. However from the reconstructed stain exclusion patterns it is often difficult to identify distinct stain excluding regions with specific protein subunits. To this end it has been demonstrated that in some cases this ambiguity can be resolved by a combination of stoichiometric labeling of the ordered structures with subunit-specific antibody fragments (e.g. Fab) and image processing of the electron micrographs recorded from labeled and unlabeled structures.


Author(s):  
Christopher Viney

Light microscopy is a convenient technique for characterizing molecular order in fluid liquid crystalline materials. Microstructures can usually be observed under the actual conditions that promote the formation of liquid crystalline phases, whether or not a solvent is required, and at temperatures that can range from the boiling point of nitrogen to 600°C. It is relatively easy to produce specimens that are sufficiently thin and flat, simply by confining a droplet between glass cover slides. Specimens do not need to be conducting, and they do not have to be maintained in a vacuum. Drybox or other controlled environmental conditions can be maintained in a sealed chamber equipped with transparent windows; some heating/ freezing stages can be used for this purpose. It is relatively easy to construct a modified stage so that the generation and relaxation of global molecular order can be observed while specimens are being sheared, simulating flow conditions that exist during processing. Also, light only rarely affects the chemical composition or molecular weight distribution of the sample. Because little or no processing is required after collecting the sample, one can be confident that biologically derived materials will reveal many of their in vivo structural characteristics, even though microscopy is performed in vitro.


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