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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Archives of microbiology 114 (1977), S. 123-136 
    ISSN: 1432-072X
    Keywords: Fungus ; Cytochemistry ; Microbodies ; Development ; Entophlyctis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Ultrastructural localization of peroxidatic activity was investigated in the chytrid Entophlyctis variabilis with the 3,3′-diaminobenzidine (DAB) cytochemical prodedure. The subcellular distribution of reaction product varied with changes in pH of the DAB medium and with the developmental stage of the fungus. Incubations in the DAB reaction medium at pH 9.2 produced an electron dense reaction product within single membrane bounded organelles which resembled microbodies but which varied in shapes from elongate to oval. At this pH the cell wall also stained darkly. When the pH of the DAB medium was lowered to pH 8.2 or 7.0, DAB oxidation product was localized within mitochondrial cristae as well as in microbodies and zoosporangial walls. As soon as zoospores were completely cleaved out of the zoosporangial cytoplasm, endoplasmic reticulum (ER) also stained. When the wall appeared around the encysted zoospore, ER staining was no longer found. The influence of the catalase inhibitor, aminotriazole, and the inhibitors of heme enzymes, sodium azide and sodium cyanide, on the staining patterns within cells incubated in the DAB media indicates that microbody staining is due to both catalase and peroxidase, mitochondrial staining is due to cytochrome c, and ER staining is due to peroxidase.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Archives of microbiology 111 (1976), S. 59-71 
    ISSN: 1432-072X
    Keywords: Fungus ; Microscopy ; Discharge apparatus ; Morphogenesis ; Extracellular material ; Taxonomy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Correlative light and electron microscopic observations were used to reconstruct the morphological events involved in the development of the discharge apparatus of Entophlyctis zoosporangia. A discharge plug formed as vesicles containing fibrillar material fused with the plasma membrane and deposited their matrices between the plasma membrane and zoosporangial wall. At the apex of the enlarging plug, the zoosporangial wall lost its microfibrillar appearance, became diffuse, and left an inoperculate discharge pore. The discharge plug exuded through this pore and then expanded into a sphere which rested at the tip of the discharge papilla or tube. After the release of the discharge plug, the number of fibrilla containing vesicles decreased and abundant endoplasmic reticulum appeared in the cytoplasm below the plug. Granular material then accumulated at the interface of the discharge plug and the plasma membrane. This was the endo-operculum. A single layer of endoplasmic reticulum subtended the area of plasma membrane which the endo-operculum covered. Later, dictyosomes appeared in the cytoplasm below the endo-operculum. Fusion of Golgi vesicles with the plasma membrane below the endo-operculum coincided with the initiation of cytoplasmic cleavage. This sequence of events indicates that, unlike the discharge plug, the endo-operculum does not originate by vesicular addition of preformed material.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Protoplasma 98 (1979), S. 177-198 
    ISSN: 1615-6102
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The ultrastructure of microbodies in developing zoosporangia ofEntophlyctis variabilis was studied by three dimensional reconstructions from serial sections and by cytochemical localization of catalase activity. The morphology of microbodies and the spatial association of microbodies with other organelles varied during fungal development. In incipient zoo-sporangia, granular dilations resembling microbodies arose from rough ER. Young, enlarging zoosporangia contained elongate, contorted microbodies continuous with ER and aligned along bundles of microtubules. Oval, paired microbodies, lying on each side of an ER cisternae, were found in all zoosporangia, but in older zoosporangia this configuration of microbodies predominated. Analysis of serial sections revealed that these oval, paired microbodies were sometimes continuous with each other, with ER, and also apparently with the ER cisterna interposed between them. Other paired, oval microbodies were clearly discrete. Constrictions were found along the length of elongate microbodies and at junctions between oval microbodies. These constrictions may represent stages in fragmentation of microbodies from pre-existing microbodies. These observations suggest that microbodies originated in three ways: 1. as local dilations in tubular ER, 2. as lateral buds from opposite sides of ER cisternae, and 3. as fragments from elongate microbodies. Microbodies were consistently spatially associated with ER, nuclear envelopes, and mitochondria. The cisterna of ER passing between paired microbodies sometimes extended into a branching, tubular system of ER which curved around the side of one microbody and lay between this microbody and the forming face of a dictyosome. The cytochemical localization of thiamine pyrophosphatase activity in this cisterna when it is not associated with dictyosomes suggests a role in metabolic control. These spatial associations indicate that the microbody assemblage with other organelles represents functional units where propinquity to other organelles and intraluminal continuities insure a system for transport of substrates and products.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1615-6102
    Keywords: Fungi ; Gilbertella persicaria ; Membranes ; Mitochondria ; Organelle isolation ; Plasma membrane ; Ultrastructure ; Vacuoles
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Methods are described for isolating and identifying subcellular membranes from walled hyphae ofGilbertella persicaria. Differences in thickness and symmetry of membranes and in contents of vesicles were used to distinguish different types of membranes. Mitochondria, vacuoles, plasma membrane, and vesicles with attached ribosomes from homogenized germlings equilibrated at the 1.2/1.4 M interface in discontinuous sucrose gradients. Accelerated flotation in centrifuged Ficol-sucrose gradients resulted in the additional separation of the mixed membranes into three fractions: one contained predominantly intact mitochondria, another was composed of vacuoles and vesicles coated with ribosomes, and a third was enriched in plasma membranes. Based upon morphometric analysis, these fractions contained 92% mitochondria, 53% vacuoles, and 89% plasma membranes, respectively. The source of vesicles coated with ribosomes was investigated since rapidly growing hyphae ofG. persicaria contained little rough endoplasmic reticulum as compared with other classes of membranes. Reconstruction from electron micrographs of mitochondrial fragmentation and vesiculation suggested that most of the ribosome-coated vesicles originated from disrupted mitochondria rather than from rough endoplasmic reticulum. The study demonstrates the utility of ultrastructural markers to identify membranesin vitro independent of, or as an adjunct to, cytochemical and biochemical markers.
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Protoplasma 175 (1993), S. 161-172 
    ISSN: 1615-6102
    Keywords: Carbohydrates ; Cell surface ; Lectins ; Oomycetes ; Saprolegnia ; Spores
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary In asexual reproduction of the water mold,Saprolegnia ferax, four distinct and sequentially produced spores are involved in dispersal, two of which are motile and two of which are nonmotile. Composition of cell surface glycoproteins may be important in dispersal strategies for each of these stages. Binding patterns of fluorescently labelled lectins were investigated to identify differences in glycoproteins of asexually produced dispersal stages. The pattern of lectin binding to zoospores was diverse. FITC-Con A bound to surfaces of zoospores and membranes of the water expulsion vacuole system, indicating the prescence of mannosyl and glucosyl residues. In zoospores incubated for more than 30 min in FITC-WGA and FITC-GS II. which bind N-acetyl glucosamine, fluorescence was sometimes localized in peripheral, intracellular patches. In shorter incubations, secondary zoospores bound these lectins along the groove region where K-bodies were located. Surfaces of cystospores typically bound FITC-WGA, but not FITC-GS II. FITC-GS II, however, bound to empty cystospore walls, probably because reactive sugars were available at the inner surface of the wall. Germ tubes emerging from cystospores bound labelled WGA and GS II, but not Con A. The same lectin binding pattern was found along discharge papilla of primary cystospores, indicating that modifications in cystospore walls associated with direct germination and zoospore discharge were similar. Thus, glycoproteins involved in early establishment of the hyphal system differ from those forming the cell surface of cystospores. Differences in the binding pattern of lectins to zoospores and cystospores highlight differences between cell surface carbohydrates of motile and nonmotile asexual stages.
    Type of Medium: Electronic Resource
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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    Protoplasma 181 (1994), S. 123-141 
    ISSN: 1615-6102
    Keywords: Carbohydrates ; Chytridiomycetes ; Extracellular material ; Membranes ; Ultrastructure ; Zoospores
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary In development of the primitive fungi, chytridiomycetes, unwalled zoospores bearing single, posterior flagella are transformed into walled, round-cells which elaborate the thallus. Production, structural modification, or release of extracellular material are involved with each transition of developmental stage. This article reviews the variety and developmental changes of extracellular materials found at the cell surface of chytridiomycetes. A cell coat, produced from Golgi-derived vesicles during zoosporogenesis, is visible around free swimming zoospores of some chytridiomycetes. How the zoospore surface receives and transduces signals is not widely explored, but it is known that fenestrated cisternae and simple cisternae, which are integrated into the microbody-lipid globule complex, are spatially and structurally associated with the plasma membrane and flagellar apparatus. This spatial association, as well as the cytochemical localization of calcium in fenestrated cisternae, suggest a mechanism for signal transduction and for regulation of zoospore motility. Zoospores become encased in a new layer of extracellular material as the zoospore encysts. Among some chytrids the source of this material is preexisting vesicles which fuse with the plasma membrane. Among other zoospores, a readily identifiable population of encystment vesicles is not apparent, demonstrating that there is no single pattern or mechanism for zoospore encystment in chytridiomycetes. Encysted zoospores developing into thalli, typically produce cell walls with a microfibrillar substructure. Ultrastructural analysis of walls reveals distinctive architecture and remarkable sculpturing which have been used in systematics of some members of chytridiomycetes. Nothing is known as to underlying controls of cytoskeletal elements and plasma membrane enzyme complexes in wall biogenesis. Many changes in cell surface structures accompany thallus maturation. Septa, many traversed with plasmodesmata, are produced in most chytrid thallus types. As sporangia and resting spores prepare for the production and release of zoospores, additional extracellular layers of material are frequently produced. Polarized deposits of extracellular material become discharge plugs, discharge vesicles, or endoopercula. Interstitial material is also released into cleavage furrows. Circumscissile or localized digestion of walls produce operculate or inoperculate exit ports for zoospore release. Cryofixation preserves more extensive extracellular material than does conventional chemical fixation, and broader application of cryofixation may radically alter our current view of cell surface structure. Thus chytridiomycetes exhibit a range in patterns for the occurrence and subsequent modifications of extracellular materials, even for members within the same order. The most universally recognized role for these extracellular materials is protection. Although there is a reasonable view of the types of extracellular material involved in chytridiomycete development, we have only limited understandings of their biogenesis or roles in regulation and communication, areas awaiting more investigations.
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 1615-6102
    Keywords: Fungus ; Zoospore ; Ultrastructure ; Membranes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Determining how the orientation and association among organelles are maintained within zoospores of theChytridiales is important to understanding the control of zoospore motility. Zoospores of the aquatic fungi,Chytriomyces aureus andC. hyalinus, contain microbody-lipid globule complexes with an elongate microbody adjacent to the portion of a lipid globule facing the cell's interior and a fenestrated cisterna (the rumposome) opposed to the surface of the lipid globule toward the plasma membrane. Mitochondria are intimately associated with the microbody. Electron microscopy of the microbody-lipid globule complex fixed in glutaraldehyde and osmium tetroxide, with or without tannic acid, reveals cross-linking bridges connecting the rumposome to the plasma membrane, to the microbody, and to microtubules of the rootlet extending from the kinetosome. It is concluded that these bridges are responsible, at least in part, for the consistent location of the microbody-lipid globule complex in the zoospore body. The possible role of the rumposome as a receptor organelle is discussed.
    Type of Medium: Electronic Resource
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  • 8
    ISSN: 1615-6102
    Keywords: Adhesion ; Carbohydrates ; Exocytosis ; K-bodies ; Lectins ; Saprolegnia ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Electron and fluorescence microscopy were used to identify organelles involved in attachment of secondary zoospores ofSaprolegnia ferax as they were transformed into secondary cysts. When secondary zoospores were exposed to 1.0% peptone in the absence or presence of a substrate, they began to encyst. If substrates were present when encystment was induced, the groove surface of the secondary zoospores adhered to them. The first event in attachment was secretion of contents of the kinetosome-associated organelle (K-body), which was typically oriented with the tubule-filled cavity positioned toward the cell surface of the groove region in the zoospore. The tubules which contained carbohydrates became coarsely granular, the matrix became more fibrous, and the shell remained along the membrane concavity that was formed as the K-body fused with the plasma membrane. Five minutes later, a cyst coat appeared, and cysts were not readily dislodged from a substrate. The concavity was no longer found, presumably because it had evaginated; but a layered pad of adhesion material was between the cyst coat and substrate. The layers of the adhesion pad corresponded to the structure of the matrix of K-bodies. As with the tubules of the K-body, the coarsely granular portion at the edge of the pad stained for carbohydrates. Similarly, the lectins WGA and GS-II labeled with fluorescein stained the rim of the adhesion pad on cysts, indicating the presence of glycoconjugates containing N-acetylglucosamines. Because globular areas near the kinetosomes and groove of zoospores (where K-bodies were located) also bound WGA and GS-II, K-bodies contained the same carbohydrates as the adhesion pad. We conclude that K-bodies function in the attachment of encysting zoospores to substrates as the cell differentiates. The tubular portion of the K-body matrix contains carbohydrates which might assist in the adhesion process.
    Type of Medium: Electronic Resource
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  • 9
    ISSN: 1615-6102
    Keywords: Protists ; Algae ; Fungi ; Protozoa ; Cell surface structures ; Terminology ; Nomenclature
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The use of a precise terminology is important to the unambiguous exchange of information in the multidisciplinary area of protistology. In this paper we attempt to establish clear definitions, give illustrations, and comment on the different terms used for cell surface structures of protists and related organisms.
    Type of Medium: Electronic Resource
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  • 10
    Electronic Resource
    Electronic Resource
    Springer
    Protoplasma 89 (1976), S. 1-27 
    ISSN: 1615-6102
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A correlative approach, involving light and electron microscopic, cytochemical, and biochemical techniques, was used to study the structure and function of microbodies in zoospores ofEntophlyctis sp. The same population of microbodies already existing in the zoosporangium appeared to be segregated into zoospore initials during cytoplasmic cleavage. Microbodies laid at the anterior end of zoospores and were part of an organized assemblage of organelles, the microbody-lipid globule complex. In the microbody-lipid globule complex, endoplasmic reticulum occurred on the surface of the lipid globules toward the zoospore's exterior, and the microbody, subtended by mitochondria, was appressed to the opposite surface of the lipid globule. The organization of the microbody-lipid globule complex changed as the zoospore swam and encysted. As lipid globules coalesced, the microbody-lipid globule complex became disorganized. After lipid globule coalescence was completed, the microbody-lipid globule complex regained its order, and several microbodies were clustered adjacent to a single lipid globule. The microbodies persisted even in the encysted zoospore, but they were found on all sides of the lipid globule. Microbodies isolated from zoospores contained catalase as well as malate synthase and isocitrate lyase, two enzymes of the glyoxylate cycle. When zoospores encysted greater activities of these glyoxylate cycle enzymes could be detected. The presence of glyoxylate cycle enzymes and the close association between the microbody and lipid globule suggest that microbodies function as glyoxysomes in zoospores and encysted zoospores. The functional significance of the morphological organization of the microbody-lipid complex is discussed in terms of energy production and the conversion of storage lipid into structural components of the cell.
    Type of Medium: Electronic Resource
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