Ultrastructure and lipid identification during conidium germination of Stemphylium sarcinaeforme

1976 ◽  
Vol 22 (1) ◽  
pp. 92-100 ◽  
Author(s):  
Gordon M. Murray ◽  
Douglas P. Maxwell

Multicelled conidia of Stemphylium sarcinaeforme germinate in water forming several germ tubes. Individual cells within conidia are connected by pores which are plugged in ungerminated conidia and open in germinated ones. During germination, vacuoles enlarge, endoplasmic reticulum profiles increase in number, and mitochondria change from spherical to elongate. The germ tube wall is laid down at the site of emergence from the conidium. Shortly after germination, a septum with a central pore forms where the germ tube emerged. The germ tube wall is surrounded by a fibrillar sheath. Lipid bodies are closely associated with vacuoles during germination. The ultrastructural location of lipid was found by extraction of conidia with lipid solvents. Total lipid decreases from 14.4% of the dry weight of ungerminated conidia to 13.4% of the dry weight of conidia germinated for 10 h. No qualitative changes occurred in the major lipid classes of conidia during germination. The activities of lipase and acid phosphatase were detected in ungerminated and germinated conidia.


1978 ◽  
Vol 56 (22) ◽  
pp. 2865-2872 ◽  
Author(s):  
Ichiko Tsuneda ◽  
Lorene L. Kennedy

Germination of basidiospores in Fomes fomentarius (Fries) Kickx is bipolar with germ tubes emerging at both ends. Ungerminated spores are smooth with a thick cell wall consisting of two layers: an outer thin, electron-dense layer and an inner thick, electron-light layer. During the early stage of germination, two additional cell wall layers are formed: a very thin, electron-dense layer and a relatively thick, electron-light layer. Germ tube walls originate from these newly formed, inner layers. Ungerminated spores are uninucleate and contain numerous lipid bodies, ribosomes, and cisternae of endoplasmic reticulum. Germinated spores have distinct mitochondria and an invaginated plasma membrane and are usually devoid of endoplasmic reticulum.



1974 ◽  
Vol 52 (11) ◽  
pp. 2335-2340 ◽  
Author(s):  
Gordon M. Murray ◽  
Douglas P. Maxwell

Multiseptate conidia of Cochliobolus carbonus germinate in water by one or two polar germ tubes. The septa between cells within a conidium contain a central pore, which is plugged before germination but open during germination, so that all cells are interconnected. During germination, vacuoles enlarge, endoplasmic reticulum profiles increase in number, and mitochondria change from a spherical form with loosely arranged cristae to an elongated form with closely packed cristae. The conidium wall consists of an inner electron-translucent layer and an outer electron-opaque layer. The germ tube wall is continuous with the innermost layer of the conidium wall. Shortly after germination, a septum with a central pore forms in the germ tube by invagination where it emerges from the conidium. The germ tube wall is surrounded by a fibrillar sheath. The presence of apical vesicles in the germ tube tip is confirmed. The associations of Woronin bodies with septal pores and microbodies, of lipids with vacuoles, and of microbodies with lipids and vacuoles are discussed.



1973 ◽  
Vol 19 (8) ◽  
pp. 1031-1034 ◽  
Author(s):  
S. D. Steele ◽  
T. W. Fraser

The dormant arthrospore in Geotrichum candidum has three, possibly four, layers making up the spore wall. Nuclei, mitochondria, free ribosomes, fragments of endoplasmic reticulum, various small vacuoles, and particles of glycogen were observed within the protoplasm. During germination a new layer of wall material forms between the original spore wall and the cytoplasm. This new layer is confined to the region where germ-tube emergence occurs and is continuous with the germ-tube wall. After germ-tube emergence vesicles were seen at the apices of germlings. Another feature of germination was an apparent increase in the amount of endoplasmic reticulum, some of which appears to assume the function of the Golgi apparatus.



1983 ◽  
Vol 29 (11) ◽  
pp. 1514-1525 ◽  
Author(s):  
Patrick A. Sullivan ◽  
Chiew Yoke Yin ◽  
Christopher Molloy ◽  
Matthew D. Templeton ◽  
Maxwell G. Shepherd

The uptake of nutrients (glucose, glutamine, and N-acetylglucosamine), the intracellular concentrations of metabolites (glucose-6-phosphate, cyclic AMP, amino acids, trehalose, and glycogen) and cell wall composition were studied in Candida albicans. These analyses were carried out with exponential-phase, stationary-phase, and starved yeast cells, and during germ-tube formation. Germ tubes formed during a 3-h incubation of starved yeast cells (0.8 × 108 cells/mL) at 37 °C during which time the nutrients glucose plus glutamine or N-acetylglucosamine (2.5 mM of each) were completely utilized. Control incubations with these nutrients at 28 °C did not form germ tubes. Uptake of N-acetylglucosamine and glutamine was inhibited by cycloheximide which suggests that de novo protein synthesis was required for the induction of these uptake systems. The glucose-6-phosphate content varied from 0.4 nmol/mg dry weight for starved cells to 2–3 nmol/mg dry weight for growing yeast cells and germ tube forming cells. Trehalose content varied from 85 nmol/mg dry weight (growing yeast cells and germ tube forming cells) to 165 nmol/mg weight (stationary-phase cells). The glycogen content decreased during germ-tube formation (from 800 to 600 nmol glucose equivalent/mg dry weight) but increased (to 1000 nmol glucose equivalent/mg dry weight) in the control incubation of yeast cells. Cyclic AMP remained constant throughout germ-tube formation at 4–6 pmol/mg dry weight. The total amino acid pool was similar in exponential, starved, and germ tube forming cells but there were changes in the amounts of individual amino acids. The overall cell wall composition of yeast cells and germ tube forming cells were similar: lipid (2%, w/w); protein (3–6%), and carbohydrate (77–85%). The total carbohydrates were accounted for as the following fractions: alkali-soluble glucan (3–8%), mannan (20–23%), acid-soluble glucan (24–27%), and acid-insoluble glucan (18–26%). The relative amounts of the alkali-soluble and insoluble glucan changed during starvation of yeast cells, reinitiation of yeast-phase growth, and germ-tube formation. Analysis of the insoluble glucan fraction from cells labelled with [14C]glucose during germ-tube formation showed that the chitin content of the cell wall increased from 0.6% to 2.7% (w/w).



1980 ◽  
Vol 26 (1) ◽  
pp. 21-26 ◽  
Author(s):  
M. G. Shepherd ◽  
Chiew Yoke Yin ◽  
S. P. Ram ◽  
P. A. Sullivan

A reproducible and simple system for the production of germ tubes from yeast cells of Candida albicans using glucose and glutamine as substrates has been described.During germ tube formation there was a doubling of the dry weight but the number of cells remained constant. Although the DNA content did not change for the first 4 h of germ tube formation, the RNA content more than doubled. The DNA and RNA content of C. albicans blastospores are 4.5 × 10−15 g per cell and 48 × 10−15 g per cell respectively.Nystatin, phenethyl alcohol, 2,4-dinitrophenol, azaserine, salicylhydroxamic acid, and 5-fluorocytosine were all effective inhibitors of germ tube formation. Cysteine, potassium cyanide, and polyoxin D did not prevent germination. The incorporation of both uracil and leucine occurred rapidly during germ tube formation. The inhibitors of RNA synthesis, actinomycin D, cordycepin, and daunomycin prevented germination and inhibited uracil incorporation. The translational inhibitors, trichodermin, aurin tricarboxylic acid, puromycin, and cyloheximide were effective in inhibiting both germ tube formation and leucine incorporation.



1970 ◽  
Vol 16 (5) ◽  
pp. 273-280 ◽  
Author(s):  
N. L. Mitchell ◽  
W. E. McKeen

Measurements made from electron micrographs of serial sections and from thoroughly plasmolyzed conidia indicate that more than 50% of the volume of the conidia of Sphaerotheca macularis consists of vacuoles in which most of the water in the conidia is stored. Electron-dense granules inside the vacuoles evidently include storage materials. Some developing vacuoles, particularly those of the germ tube, enclose membrane-bound bodies resembling lysosomes which later disappear as the vacuoles enlarge. Conspicuous multimembraned myelin-like bodies project inside the vacuolar cavity, their membranes being continuous with the vacuolar membrane. These bodies are believed to function in the synthesis of new cytoplasmic materials from the reserves in the vacuoles.The conidium, which may later produce up to four germ tubes, always retains a nucleus. The nucleus contains a peripheral granule which maintains a leading position on migrating nuclei and divides into two during the initial stages of nuclear division.Germ tubes respond positively to the stimulus of unilateral illumination and are produced on the illuminated sides of the conidia. Cytoplasmic changes which accompany germination include the increase in number and size of mitochondria, particularly in the germ tube. Their multiplication appears to be by fission. Endoplasmic reticulum is greatly increased and ribosomes are more abundant. Aggregated granules resembling glycogen particles also occur, these not being usually seen in resting conidia.



1991 ◽  
Vol 69 (11) ◽  
pp. 2455-2467 ◽  
Author(s):  
C. Gerald Van Dyke ◽  
Charles W. Mims

Nongerminating conidia of Colletotrichum truncatum were coated with copious amounts of a finely fibrillar extracellular matrix. This matrix spread out onto the dialysis membrane used as a substrate in this study. Each thin-walled conidium contained a single nucleus that underwent mitosis 1–2 h following placement of aqueous suspensions of conidia on membranes. A septum subsequently developed near the middle of the conidium, creating two uninucleate cells. Just prior to or during septum development a germ tube emerged laterally, usually near one end of the conidium. The nucleus moved into the germ tube and underwent mitosis. One daughter nucleus remained in the germ tube, the other moved back into the conidium. Developing germ tubes appeared to produce large amounts of electron-dense, fibrillar material that coated their surfaces. This material blended into the remnants of the matrix initially coating conidia and could not be clearly differentiated from the latter material. Germ tubes grew to various lengths before forming appressoria. Appressorium differentiation began shortly after the germ-tube tip curved sharply. A septum developed to delimit the tip that differentiated into a swollen appressorium. By 6 h following initial hydration of conidia, appressoria were melanized and the surrounding extracellular material had condensed onto their surfaces, forming an electron-dense coating that appeared to stick appressoria to dialysis membranes. A tiny penetration peg developed from an apparently wall-less region on the underside of the mature appressorium and, in some instances, grew a short distance into the dialysis membrane. Key words: electron microscopy, freeze substitution, conidia, appressoria.



2006 ◽  
Vol 52 (2) ◽  
pp. 117-124 ◽  
Author(s):  
Ariani Corrêa Barbosa ◽  
Anousca Evelyn do Carmo ◽  
Letícia Graf ◽  
Roberto Tomaz ◽  
Caroline Fogaça de Souza ◽  
...  

Colletotrichum acutatum may develop one or more secondary conidia after conidial germination and before mycelial growth. Secondary conidia formation and germination were influenced by conidia concentration. Concentrations greater than 1 × 105conidia/mL were associated with germination decrease and with secondary conidia emergence. Secondary conidia can form either alone or simultaneously with germ tubes and appressoria. Confocal analysis showed numerous lipid bodies stored inside ungerminated conidia, which diminished during germ tube and appressoria formation, with or without secondary conidia formation. They were also reduced during secondary conidia formation alone. While there was a decrease inside germinated conidia, lipid bodies appeared inside secondary conidia since the initial stages. Intense vacuolization inside primary germinated conidia occurred at the same time as the decrease in lipid bodies, which were internalized and digested by vacuoles. During these events, small acidic vesicles inside secondary conidia were formed. Considering that the conidia were maintained in distilled water, with no exogenous nutrients, it is clear that ungerminated conidia contain enough stored lipids to form germ tubes, appressoria, and the additional secondary conidia replete with lipid reserves. These results suggested a very complex and well-balanced regulation that makes possible the catabolic and anabolic pathways of these lipid bodies.Key words: secondary conidia, lipid bodies, vacuoles, confocal microscopy, Colletotrichum.



2002 ◽  
Vol 80 (10) ◽  
pp. 1121-1125 ◽  
Author(s):  
H H Edwards

Development of primary germ tubes from conidia of Blumeria graminis f.sp. hordei on primary leaf segments of Hordeum vulgare was investigated from 3 to 13 h postinoculation (hpi) using transmission electron microscopy. By 3 hpi, the primary germ tube wall that makes contact with the host cuticle develops a small protrusion that breaches the host cuticle and touches the host cell wall but does not penetrate any further. This protrusion is the cuticular peg. From 3 to 13 hpi, the cuticular peg swells, becomes quite electron dense, and finally develops a loose fibrillar texture. The structure of the primary germ tube with the terminal cuticular peg is consistent with the hypothesis that it allows the conidium to absorb water and solutes present in the host cell wall.Key words: powdery mildew, barley, ultrastructure.



1977 ◽  
Vol 25 (5) ◽  
pp. 319-328 ◽  
Author(s):  
E Dannen ◽  
M E Beard

Organelles with the morphologic characteristics of peroxisomes have been found in the cells of the kidney sac of two terrestrial pulmonate gastropods. Arion ater and Ariolimax columbianus. These peroxisomes appear in profile as circles or ellipses, 0.25 micron in diameter and 0.3-0.8 micron long; They have a finely granular matrix and a single-limiting membrane; the organelles are extensively associated with smooth endoplasmic reticulum. Some Ariolimax peroxisomes contained structures reminiscent of nucleoids while those of Arion did not. The peroxisomes of Arion ater show a strongly-positive staining reaction with the 3,3'-diaminobenzidine technique, which is inhibited in the presence of aminotriazole. Peroxisomes of Ariolimax columbianus did not show a positive reaction, despite a number of variations of the 3,3'-diaminobenzidine protocol. Speculations are made concerning the biochemical reasons for this cytochemical behavior. Peroxisomes in both tissues were negatively stained while lysosomes were positively stained in acid-phosphatase incubations.



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