Fine structure of the macroconidia of Fusarium solani

1975 ◽  
Vol 53 (19) ◽  
pp. 2134-2146 ◽  
Author(s):  
J. P. Tewari ◽  
W. P. Skoropad
Keyword(s):  
Cell Wall ◽  
Fusarium Solani ◽  
Superficial Layer ◽  
Dense Layer ◽  
Inoculum Potential ◽  
Chemical Fixation ◽  
Thin Sections ◽  
Innermost Layer ◽  
Freeze Etching ◽  
Myelin Figures

Ultrastructure of the macroconidia of Fusarium solani as visualized by transmission (ultrathin sectioning and freeze-etching) and scanning electron microscopy is described. The cell wall has four layers. The innermost layer is electron-lucid followed by an electron-dense layer. The next outer layer is spongy in appearance followed by a superficial layer consisting of fine filamentous processes. Freeze-etch replicas of conidia directly removed from the sporodochia and still suspended in the mucilaginous material (in which they are produced) frequently show the conidia connected by the superficial filamentous processes in the cell wall. This agglutination of the conidia is likely to increase the inoculum potential of this pathogen at the sites of infection. Structure of various membrane systems in the cells is described. The endoplasmic reticulum is fairly extensive and fenestrated. Thin sections of routinely fixed conidia show myelin figures. However, such structures were not seen in replicas of conidia that were freeze-etched without use of chemical fixation or cryoprotection.

Ultrastructure of Staphylococcus epidermidis after freeze-etching and thin sectioning

1973 ◽  
Vol 19 (2) ◽  
pp. 294-295
Author(s):  
James E. Gilchrist ◽  
Irving W. DeVoe
Keyword(s):  
Staphylococcus Aureus ◽  
Outer Layer ◽  
Staphylococcus Epidermidis ◽  
Cell Walls ◽  
Cytoplasmic Membrane ◽  
Middle Layer ◽  
Dense Layer ◽  
Thin Sections ◽  
Thin Sectioning ◽  
Freeze Etching

A considerable quantity of information is now available on the ultrastructure of Staphylococcus (1, 2, 4, 7, 8, 10, 11, 12). Cell walls of these organisms in thin sections have been shown to consist of three layers: a dense outer layer, a rather electron translucent middle layer, and a very dense layer next to the cytoplasmic membrane (2, 7, 11, 12). Bulger and Bulger (2) have pointed out the presence of circumferential substructure in the middle layer of the wall in a strain of Staphylococcus aureus isolated as the causative agent in fatal pneumonia.Numerous mesosomes of both the vesicular and laminar types are evident in thin sections of staphylococci from several studies (1, 4, 7, 11). Moreover, single vesicular structures that appear to be invaginations of the trilaminar cytoplasmic membrane have been pointed out by Suganuma (11) and Beaton (1).

scholarly journals Ultrastructure of Lipid Deposits and Other Contents in Freeze-Etched Coleoptile Cells of Ungerminated Rice Grains

1973 ◽  
Vol 26 (2) ◽  
pp. 357 ◽  
Author(s):  
MS Buttrose ◽  
A Soeffky
Keyword(s):  
Aqueous Media ◽  
Chemical Fixation ◽  
Thin Sections ◽  
Rice Grains ◽  
Freeze Etching

Freeze-etching has been used to avoid exposure to aqueous media in studying ultrastructure in coleoptile cells of ungerminated rice grains. Results differ in some respects from those obtained after chemical fixation and examination of thin sections.

Mesosome-like Structures in Cryptococcus neoformans Studied by Chemical Fixation and by the Freeze-etching Technique

1968 ◽  
Vol 26 ◽  
pp. 80-81
Author(s):  
M. R. Edwards ◽  
S. C. Holt
Keyword(s):  
Electron Microscope ◽  
Cryptococcus Neoformans ◽  
Room Temperature ◽  
Osmium Tetroxide ◽  
Chemical Fixation ◽  
Etching Technique ◽  
Pathogenic Yeast ◽  
Thin Sections ◽  
Lead Salts ◽  
Freeze Etching

The general features of Cryptococcus neoformans, a pathogenic yeast, have been studied with the electron microscope. In the course of such a study it was noted that the plasma membrane of C. neoformans, occasionally invaginated into the cytoplasm and formed membranous organelles which resembled bacterial mesosomes. The present investigation was undertaken in order to examine such structures in detail and to compare the results from chemical fixation with those of freeze-etching.Cells were grown in Sabouraud's agar at 25-27° C for 24-48 hr and fixed with 4% glutaraldehyde in 0.15 M phosphate (Sbrensen's) buffer, at room temperature, for 2 hr; after being thoroughly washed in the buffer and post-fixed in osmium tetroxide, in the same buffer, they were dehydrated in ethyl alcohol and embedded in Epon. Thin sections were cut in a LKB microtome, double stained with uranyl and lead salts and examined in the Siemens Elmiskop IA.

Cytochemistry of acid mucosubstance and acid phosphatase in Cryptococcus neoformans

1974 ◽  
Vol 20 (6) ◽  
pp. 833-838 ◽  
Author(s):  
Thomas A. Mahvi ◽  
Samuel S. Spicer ◽  
Nancy J. Wright
Keyword(s):  
Cell Wall ◽  
Acid Phosphatase ◽  
Cryptococcus Neoformans ◽  
Ultrastructural Level ◽  
Dense Layer ◽  
Strong Activity ◽  
Thin Sections ◽  
Cytoplasmic Bodies ◽  
Cryostat Sections ◽  
Substrate Medium

At the ultrastructural level, dialyzed iron stains only the mucoid capsule in Cryptococcus neoformans, converting it from the most lucent to an extremely dense layer and thus demonstrating its content of acid mucosubstance. After lead staining of thin sections of dialyzed iron treated specimens, the wall exhibits a dense inner and translucent outer layer. The inner layer and mucoid capsule appear thicker in the mother than the daughter cell of budding organisms. C. neoformans cells fixed in suspension and incubated in acid phosphatase substrate medium exhibit at the ultrastructural level activity confined to the mucoid capsule, cell wall, and internal extensions presumed to be plasmalemmasomes. Organisms in small blocks or cryostat sections of pellets fixed in clotted fibrin, when incubated for acid phosphatase, reveal moderate to no reactivity in cell wall and mucoid capsule but strong activity in surrounding fibrin aggregates. In the latter specimens, reaction product indicative of acid phosphatase also is evident in the nuclear envelope, in some of the profiles of presumed granular reticulum, in complex lamellar profiles presumed to be Golgi elements, and in small cytoplasmic bodies and large vacuoles. The morphological specimens disclose counterparts of acid phosphatase reactive structures and show lucent cytoplasm segregated by circular mitochondrial profiles and a difference between the thickness and density of the plasmalemma and other cell membranes.

Superficial cell-wall layers on Spirillum "Ordal" and their in vitro reassembly

1976 ◽  
Vol 22 (4) ◽  
pp. 567-582 ◽  
Author(s):  
T. J. Beveridge ◽  
R. G. E. Murray
Keyword(s):  
Cell Envelope ◽  
Amorphous Layer ◽  
Superficial Layer ◽  
Optical Diffraction ◽  
Hexagonal System ◽  
Thin Sections ◽  
Regular Structures ◽  
Packed Layer ◽  
Freeze Etching

The cell envelope of Spirillum sp. strain "Ordal" (possibly a variety of S. anulus) demonstrated multiple superficial wall layers which were diverse in their macromolecular arrays. Negative staining and freeze-etching techniques revealed an outer hexagonally packed layer and an inner tetragonally packed layer. However, both thin sections and freeze-etched cleavages of the wall showed that each of these regular structures rested upon a backing layer, and that there was a delicate amorphous layer overlying the outer hexagonal array. Rotary integration, optical diffraction, and reconstruction of image were used to clarify measurements of each array and to verify the validity of a diagrammatic model of the outer hexagonal system. The integrity of these layers required suitable cations (Ca2+ appeared essential) and pH ([Formula: see text] dissociated most superficial layers). These observations aided in the development of a low-pH cationic-substitution technique, in which Na+ replaced essential Ca2+, for extraction of the layers from the cell surface. Dialysis to remove Na+ and restoration of Ca2+ initiated in vitro reassembly of the superficial layer components until regularly structured assembly products were formed.

Ultrastructural changes in the conidia of Penicillium notatum during germination

1973 ◽  
Vol 19 (7) ◽  
pp. 797-801 ◽  
Author(s):  
J. F. Martin ◽  
F. Uruburu ◽  
J. R. Villanueva
Keyword(s):  
Cell Wall ◽  
Cell Walls ◽  
Ultrastructural Changes ◽  
Dense Layer ◽  
Penicillium Notatum ◽  
Thin Sections ◽  
Resting Spores ◽  
Different Types ◽  
Germ Tubes ◽  
Isolated Cell

To study the changes in the cell wall of Penicillium notatum during germination, thin sections of resting, swollen, and germinating spores, and mycelium were compared with thin sections of the isolated cell walls. In the cell wall of resting spores four distinct layers were found. The outermost layer of the cell wall of resting spores was released during swelling and the two inner layers were extended to form the cell wall of the germ tube. The cell wall of young germ tubes had only two layers but a new electron-dense layer was formed later on the outside. Mycelial cell walls which appeared thinner than those of conidia showed three distinct layers. Large mitochondria that divide during germination were present in both resting and swollen spores. Two different types of vacuoles were found, both of which decreased in size and in number during germination. Endoplasmic reticulum was almost absent in resting spores but increased substantially during swelling.

Observations of thick and thin sections in a field-emission SEM

1994 ◽  
Vol 52 ◽  
pp. 1018-1019
Author(s):  
W. P. Wergin ◽  
S. Roy ◽  
E. F. Erbe ◽  
C. A. Murphy ◽  
C. D. Pooley
Keyword(s):  
Electron Microscope ◽  
Field Emission ◽  
Room Temperature ◽  
Osmium Tetroxide ◽  
Uranyl Acetate ◽  
Chemical Fixation ◽  
Thin Sections ◽  
Transmission Electron ◽  
Conventional Transmission Electron Microscope ◽  
Scanning Electron

Larvae of the nematode, Steinernema carpocapsae Weiser strain All, were cryofixed and freezesubstituted for 3 days in acetone containing 2% osmium tetroxide according to established procedures. Following chemical fixation, the nematodes were brought to room temperature, embedded in Spurr's medium and sectioned for observation with a Hitachi S-4100 field emission scanning electron microscope that was equipped with an Oxford CT 1500 Cryotrans System. Thin sections, about 80 nm thick, similar to those generally used in conventional transmission electron microscope (TEM) studies were mounted on copper grids and stained with uranyl acetate for 30 min and lead citrate for 5 min. Sections about 2 μm thick were also mounted and stained in a similar fashion. The grids were mounted on an Oxford grid holder, inserted into the microscope and onto a cryostage that was operated at ambient temperature. Thick and thin sections of the larvae were evaluated and photographed in the SEM at different accelerating voltages. Figs. 4 and 5 have undergone contrast conversion so that the images would resemble transmitted electron micrographs obtained with a TEM.

Freeze substitution fixation of fungal reproductive structures and spores

1989 ◽  
Vol 47 ◽  
pp. 990-991
Author(s):  
C. W. Mims ◽  
E. A. Richardson
Keyword(s):  
Plant Pathogens ◽  
Freeze Substitution ◽  
Uranyl Acetate ◽  
Chemical Fixation ◽  
Dialysis Membrane ◽  
Razor Blade ◽  
Thin Sections ◽  
Reproductive Structures ◽  
Dry Down ◽  
Diamond Knife

The advantages of freeze substitution fixation over conventional chemical fixation for preservation of ultrastructural details in fungi have been discussed by various authors. As most ascomycetes, basidiomycetes and deuteromycetes do not fix well using conventional chemical fixation protocols, freeze substitution has attracted the attention of many individuals interested in fungal ultrastructure. Thus far most workers using this technique on fungi have concentrated on thin walled somatic hyphae. However, in our laboratory we have experimented with the use of freeze substitution on a variety of fungal reproductive structures and spores with promising results.Here we present data on freeze substituted samples of sporangia of the zygomycete Umbellopsis vinacea, basidia of Exobasidium camelliae var. gracilis, developing teliospores of the smut Sporisorium sorghi, germinating teliospores of the rust Gymnosporangium clavipes, germinating conidia of the deuteromycete Cercosporidium personatum, and developing ascospores of Ascodesmis nigricans.Spores of G. clavipes and C. personatum were deposited on moist pieces of sterile dialysis membrane where they hydrated and germinated. Asci of A. nigricans developed on pieces of dialysis membrane lying on nutrient agar plates. U. vinacea was cultured on small pieces of agar-coated wire. In the plant pathogens E. camelliae var. gracilis and S. sorghi, a razor blade was used to remove smal1 pieces of infected host issue. All samples were plunged directly into liquid propane and processed for study according to Hoch.l Samples on dialysis membrane were flat embedded. Serial thin sections were cut using a diamond knife, collected on slot grids, and allowed to dry down onto Formvar coated aluminum racks. Sections were post stained with uranyl acetate and lead citrate.

New insights on Laminaria digitata ultrastructure through combined conventional chemical fixation and cryofixation

2021 ◽  
Vol 0 (0) ◽  
Author(s):  
Christos Katsaros ◽  
Sophie Le Panse ◽  
Gillian Milne ◽  
Carl J. Carrano ◽  
Frithjof Christian Küpper
Keyword(s):  
Cell Wall ◽  
General Structure ◽  
Raw Material ◽  
Rocky Shores ◽  
Chemical Fixation ◽  
Laminaria Digitata ◽  
Vegetative Cells ◽  
Biological Studies ◽  
New Findings ◽  
Wide Range

Abstract The objective of the present study is to examine the fine structure of vegetative cells of Laminaria digitata using both chemical fixation and cryofixation. Laminaria digitata was chosen due to its importance as a model organism in a wide range of biological studies, as a keystone species on rocky shores of the North Atlantic, its use of iodide as a unique inorganic antioxidant, and its significance as a raw material for the production of alginate. Details of the fine structural features of vegetative cells are described, with particular emphasis on the differences between the two methods used, i.e. conventional chemical fixation and freeze-fixation. The general structure of the cells was similar to that already described, with minor differences between the different cell types. An intense activity of the Golgi system was found associated with the thick external cell wall, with large dictyosomes from which numerous vesicles and cisternae are released. An interesting type of cisternae was found in the cryofixed material, which was not visible with the chemical fixation. These are elongated structures, in sections appearing tubule-like, close to the external cell wall or to young internal walls. An increased number of these structures was observed near the plasmodesmata of the pit fields. They are similar to the “flat cisternae” found associated with the forming cytokinetic diaphragm of brown algae. Their possible role is discussed. The new findings of this work underline the importance of such combined studies which reveal new data not known until now using the old conventional methods. The main conclusion of the present study is that cryofixation is the method of choice for studying Laminaria cytology by transmission electron microscopy.

scholarly journals New methods for confocal imaging of infection threads in crop and model legumes

Plant Methods ◽  
2021 ◽  
Vol 17 (1) ◽  
Author(s):  
Angus E. Rae ◽  
Vivien Rolland ◽  
Rosemary G. White ◽  
Ulrike Mathesius
Keyword(s):  
Cell Wall ◽  
Root Hair ◽  
Periodic Acid ◽  
Confocal Imaging ◽  
Wall Material ◽  
Future Research ◽  
Thin Sections ◽  
New Methods ◽  
Infection Threads ◽  
Imaging Of Infection

Abstract Background The formation of infection threads in the symbiotic infection of rhizobacteria in legumes is a unique, fascinating, and poorly understood process. Infection threads are tubes of cell wall material that transport rhizobacteria from root hair cells to developing nodules in host roots. They form in a type of reverse tip-growth from an inversion of the root hair cell wall, but the mechanism driving this growth is unknown, and the composition of the thread wall remains unclear. High resolution, 3-dimensional imaging of infection threads, and cell wall component specific labelling, would greatly aid in our understanding of the nature and development of these structures. To date, such imaging has not been done, with infection threads typically imaged by GFP-tagged rhizobia within them, or histochemically in thin sections. Results We have developed new methods of imaging infection threads using novel and traditional cell wall fluorescent labels, and laser confocal scanning microscopy. We applied a new Periodic Acid Schiff (PAS) stain using rhodamine-123 to the labelling of whole cleared infected roots of Medicago truncatula; which allowed for imaging of infection threads in greater 3D detail than had previously been achieved. By the combination of the above method and a calcofluor-white counter-stain, we also succeeded in labelling infection threads and plant cell walls separately, and have potentially discovered a way in which the infection thread matrix can be visualized. Conclusions Our methods have made the imaging and study of infection threads more effective and informative, and present exciting new opportunities for future research in the area.

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