scholarly journals Some new aspects of the pea (Pisum sativum L.) root nodule ultrastructure

2014 ◽  
Vol 65 (3-4) ◽  
pp. 221-233 ◽  
Author(s):  
Wojciech Borucki

Unequal cell divisions were observed in the meristem of pea root nodule. Since after such divisions only the bigger cells become infected then those divisions play a significant role in the formation of the three-dimensional structure of the bacteroidal tissue. In the infected cells of the young ineffective bacteroidal tissue the first host reaction to the incompatibility of the symbiotic system is the RER membranes aggregation. In effective symbiosis RER membranes form permanent sites of contact with the peribacteroidal membranes thus connecting all the symbiosoms in the cell. Possibly that ensures the synchronisation of the differentiation processes of the bacteroids and/or their simultaneous degeneration. The presence of membraneous structures in the form of rings is a characteristic feature of effective bacteroids. It is postulated that the structures are directly connected with nitrogen assimilation. Structures X and Y which are present in the bacteroids of the effective and ineffective symbiosis may be connected with the adaptation of bacterial cells to lowered oxygen pressure in bacteroidal tissue and their transformation (structures X) into bacteroids. The presence of the cytoplasm (or cytoplasmatic remnants) of the infected cells was observed in the intercellular spaces. It is sugested that it is a way, so far unknown, of the gas diffusion regulation in bacteroidal tissue.

Author(s):  
Ekaterina M. Lenchenko ◽  
◽  
Damir I. Udavliev ◽  
Inna B. Pavlova ◽  
◽  
...  

The results of morphometric and densitometric parameters biofilms are presented, effective methods of detecting uncultivated viable microorganisms isolated from a representative sample of objects of veterinary and sanitary supervision are tested and selected. Optical, luminescent and scanning electron microscopy revealed the formation of a three-dimensional structure biofilms in the form a dense network consisting of gram-negative and gram-positive bacteria, yeast cells, hyphal and pseudohyphalic forms, surrounded by an intercellular polymer matrix. The presence hyphae of microscopic fungi causes an increase in the number of cells adhered to the substrate, microcolonies were formed from bacteria and yeast cells of microscopic fungi. The pathogenesis of the syndrome of overgrowth of microorganisms is provided by the presence of various dissociative variants, the dispersion of uncultivated bacterial cells, which gain advantages in the hyperagregation of the architectonics of heterogeneous biofilms. Multilayer membranes, vesicles, cells with a defective cell wall, spheroplasts, protoplasts, L-shapes, needle-like and giant structures, and revertant cells were identified. The dynamics of changes in the viable structures microorganisms was characterized by alternating periods of decrease and increase in the intensity of biofilm formation. When detecting the viability of microorganisms in the composition biofilms, viable and non-viable cells were differentiated – a green luminescence spectrum and a red luminescence spectrum, respectively. The dissociation of the population caused an increase in the concentration of R-dissociant cells with a higher growth rate, cell lysis was detected after 48–72 h of cultivation, a change in the ratio phenotypic forms was observed – the M-dissociant was predominant. The study of the heterogeneous structure of the population, without disturbing the natural architectonics of biofilms, revealed direct correlations (r = 0,89) between morphometric (≥90 % of the field of view) and densitometric parameters (OD). The efficiency of a nutrient medium containing pancreatic hydrolyzate, mannitol, L-asparagine and glycerol was established for the repair of the cell wall, the reversal of L-forms of microorganisms.


2011 ◽  
Vol 76 (2) ◽  
pp. 109-118 ◽  
Author(s):  
Wojciech Borucki

Vacuoles play very important physiological roles in plant cells. Pea root nodules, which exhibit distinct zonation (meristematic zone and central tissue zones), may serve as a good experimental model for the investigations of vacuole development and its importance to cell and tissue functioning. Moreover, the nodule central tissue is composed of both infected and uninfected cells which play different physiological roles and differ in the level of vacuolation. Cytological observations revealed that central vacuoles of the infected cells of the effective nodules expand toward cell walls. Thus only thin layers of the cytoplasm separate each central vacuole from plasma membrane and cell wall. This finding is discussed from the viewpoint of improved exchange of solutes and water between the central vacuole and apoplast of the infected cell. Three-dimensional reconstruction of the vacuoles of infected cells within a fragment of effective nodule central tissue, showed their spatial arrangement. Possible advantages coming from the spatial arrangement of vacuoles within the central tissue are discussed. A comparative study of the central tissue (bacteroidal tissue) and meristem vacuolation of the effective and ineffective pea root nodules is also presented. Morphometric measurements revealed that the effective nodule central tissue was more vacuolated than the ineffective one. It was proved that maturation of the infected cells involves dynamic changes in their vacuolation. Having numerous fixing nitrogen bacteroids, the infected cells of effective central tissue were less vacuolated than uninfected cells. On the other hand, both infected and uninfected cells of the effective central tissue showed a much higher level of vacuolation in nitrogen-fixing zone than cells of the same type in ineffective tissue. These results indicate that vacuolation is an important factor in development and functioning of pea root nodule central tissue.


Author(s):  
M. Boublik ◽  
W. Hellmann ◽  
F. Jenkins

The present knowledge of the three-dimensional structure of ribosomes is far too limited to enable a complete understanding of the various roles which ribosomes play in protein biosynthesis. The spatial arrangement of proteins and ribonuclec acids in ribosomes can be analysed in many ways. Determination of binding sites for individual proteins on ribonuclec acid and locations of the mutual positions of proteins on the ribosome using labeling with fluorescent dyes, cross-linking reagents, neutron-diffraction or antibodies against ribosomal proteins seem to be most successful approaches. Structure and function of ribosomes can be correlated be depleting the complete ribosomes of some proteins to the functionally inactive core and by subsequent partial reconstitution in order to regain active ribosomal particles.


Author(s):  
Robert Glaeser ◽  
Thomas Bauer ◽  
David Grano

In transmission electron microscopy, the 3-dimensional structure of an object is usually obtained in one of two ways. For objects which can be included in one specimen, as for example with elements included in freeze- dried whole mounts and examined with a high voltage microscope, stereo pairs can be obtained which exhibit the 3-D structure of the element. For objects which can not be included in one specimen, the 3-D shape is obtained by reconstruction from serial sections. However, without stereo imagery, only detail which remains constant within the thickness of the section can be used in the reconstruction; consequently, the choice is between a low resolution reconstruction using a few thick sections and a better resolution reconstruction using many thin sections, generally a tedious chore. This paper describes an approach to 3-D reconstruction which uses stereo images of serial thick sections to reconstruct an object including detail which changes within the depth of an individual thick section.


Author(s):  
T.D. Pollard ◽  
P. Maupin

In this paper we review some of the contributions that electron microscopy has made to the analysis of actin and myosin from nonmuscle cells. We place particular emphasis upon the limitations of the ultrastructural techniques used to study these cytoplasmic contractile proteins, because it is not widely recognized how difficult it is to preserve these elements of the cytoplasmic matrix for electron microscopy. The structure of actin filaments is well preserved for electron microscope observation by negative staining with uranyl acetate (Figure 1). In fact, to a resolution of about 3nm the three-dimensional structure of actin filaments determined by computer image processing of electron micrographs of negatively stained specimens (Moore et al., 1970) is indistinguishable from the structure revealed by X-ray diffraction of living muscle.


Author(s):  
J.L. Williams ◽  
K. Heathcote ◽  
E.J. Greer

High Voltage Electron Microscope already offers exciting experimental possibilities to Biologists and Materials Scientists because the increased specimen thickness allows direct observation of three dimensional structure and dynamic experiments on effectively bulk specimens. This microscope is designed to give maximum accessibility and space in the specimen region for the special stages which are required. At the same time it provides an ease of operation similar to a conventional instrument.


Author(s):  
G. E. Tyson ◽  
M. J. Song

Natural populations of the brine shrimp, Artemia, may possess spirochete- infected animals in low numbers. The ultrastructure of Artemia's spirochete has been described by conventional transmission electron microscopy. In infected shrimp, spirochetal cells were abundant in the blood and also occurred intra- and extracellularly in the three organs examined, i.e. the maxillary gland (segmental excretory organ), the integument, and certain muscles The efferent-tubule region of the maxillary gland possessed a distinctive lesion comprised of a group of spirochetes, together with numerous small vesicles, situated in a cave-like indentation of the base of the tubule epithelium. in some instances the basal lamina at a lesion site was clearly discontinuous. High-voltage electron microscopy has now been used to study lesions of the efferent tubule, with the aim of understanding better their three-dimensional structure.Tissue from one maxillary gland of an infected, adult, female brine shrimp was used for HVEM study.


Author(s):  
Jerome J. Paulin

Within the past decade it has become apparent that HVEM offers the biologist a means to explore the three-dimensional structure of cells and/or organelles. Stereo-imaging of thick sections (e.g. 0.25-10 μm) not only reveals anatomical features of cellular components, but also reduces errors of interpretation associated with overlap of structures seen in thick sections. Concomitant with stereo-imaging techniques conventional serial Sectioning methods developed with thin sections have been adopted to serial thick sections (≥ 0.25 μm). Three-dimensional reconstructions of the chondriome of several species of trypanosomatid flagellates have been made from tracings of mitochondrial profiles on cellulose acetate sheets. The sheets are flooded with acetone, gluing them together, and the model sawed from the composite and redrawn.The extensive mitochondrial reticulum can be seen in consecutive thick sections of (0.25 μm thick) Crithidia fasciculata (Figs. 1-2). Profiles of the mitochondrion are distinguishable from the anterior apex of the cell (small arrow, Fig. 1) to the posterior pole (small arrow, Fig. 2).


Author(s):  
Kenneth H. Downing ◽  
Hu Meisheng ◽  
Hans-Rudolf Went ◽  
Michael A. O'Keefe

With current advances in electron microscope design, high resolution electron microscopy has become routine, and point resolutions of better than 2Å have been obtained in images of many inorganic crystals. Although this resolution is sufficient to resolve interatomic spacings, interpretation generally requires comparison of experimental images with calculations. Since the images are two-dimensional representations of projections of the full three-dimensional structure, information is invariably lost in the overlapping images of atoms at various heights. The technique of electron crystallography, in which information from several views of a crystal is combined, has been developed to obtain three-dimensional information on proteins. The resolution in images of proteins is severely limited by effects of radiation damage. In principle, atomic-resolution, 3D reconstructions should be obtainable from specimens that are resistant to damage. The most serious problem would appear to be in obtaining high-resolution images from areas that are thin enough that dynamical scattering effects can be ignored.


Author(s):  
J.S. Wall ◽  
V. Maridiyan ◽  
S. Tumminia ◽  
J. Hairifeld ◽  
M. Boublik

The high contrast in the dark-field mode of dedicated STEM, specimen deposition by the wet film technique and low radiation dose (1 e/Å2) at -160°C make it possible to obtain high resolution images of unstained freeze-dried macromolecules with minimal structural distortion. Since the image intensity is directly related to the local projected mass of the specimen it became feasible to determine the molecular mass and mass distribution within individual macromolecules and from these data to calculate the linear density (M/L) and the radii of gyration.2 This parameter (RQ), reflecting the three-dimensional structure of the macromolecular particles in solution, has been applied to monitor the conformational transitions in E. coli 16S and 23S ribosomal RNAs in solutions of various ionic strength.In spite of the differences in mass (550 kD and 1050 kD, respectively), both 16S and 23S RNA appear equally sensitive to changes in buffer conditions. In deionized water or conditions of extremely low ionic strength both appear as filamentous structures (Fig. la and 2a, respectively) possessing a major backbone with protruding branches which are more frequent and more complex in 23S RNA (Fig. 2a).


Sign in / Sign up

Export Citation Format

Share Document