Electron microscopy studies on infection of resistant (Sr6 gene) and susceptible near-isogenic wheat lines by Puccinia graminis f. sp. tritici

1974 ◽  
Vol 52 (12) ◽  
pp. 2615-2620 ◽  
Author(s):  
R. A. Skipp ◽  
D. E. Harder ◽  
D. J. Samborski

Intracellular hyphae, haustoria, and infected host cells in young stem rust infections of a wheat line containing resistance gene Sr6 were compared by electron microscopy with those of a near-isogenic susceptible line. Mesophyll cells of the resistant line which had undergone a rapid necrotic response to haustorial invasion were collapsed and lacked the thin layer of cytoplasm that typically bounded the vacuole of infected cells of the susceptible line. Instead, the lumen was filled with an electron-dense ground material of mixed cytoplasmic and vacuolar constituents containing organelles at various stages of disorganization. Haustoria within these cells were necrotic, as were their mother cells. Apart from a dense staining of mitochondrial membranes there was little evidence of damage to the infection hypha proximal to the septum of the haustorium mother cell. The cytoplasm of host cells adjacent to those showing necrosis was often diffuse and vesiculated. A deposit of electron-dense granular material was found between the wall of a necrotic cell and its plasmalemma, and a similar deposit was present inside the adjoining wall of a contiguous non-necrotic cell.These findings are discussed in relation to the inhibition of fungal growth that accompanies host cell necrosis in the resistant wheat line.

1987 ◽  
Vol 65 (8) ◽  
pp. 1586-1598 ◽  
Author(s):  
L. Shain ◽  
U. Järlfors

The infection process in four clones of eastern cottonwood susceptible or resistant to leaf rust caused by Melampsora medusae was studied by light and electron microscopy. Infection was initiated by stomatal rather than direct entry. Typical dikaryotic haustoria were observed in all clones within 1 day of inoculation. Some healthy-appearing haustoria were observed in susceptible clones throughout the duration of the study, which was terminated during the initiation of uredial production. Incompatibility was expressed differently in the two resistant clones. In clone St 75, most haustoria and invaded host cells that were observed appeared necrotic within 2 days of inoculation. Cell wall appositions appeared during this time in cells adjoining necrotic host cells. Some infected cells disintegrated within 4 days of inoculation. Affected host cells of clone St 92, on the other hand, plasmolyzed during the first 2 to 3 days after inoculation. Necrotic host cells were not observed in this clone until the 4th day after inoculation. Hyphal ramification and host plasmolysis were extensive at 6 days after inoculation.


1994 ◽  
Vol 72 (8) ◽  
pp. 1164-1170 ◽  
Author(s):  
Theresa A. S. Aveling ◽  
Heidi G. Snyman ◽  
F. H. J. Rijkenberg

Conidial germination of Alternaria porri, formation of prepenetration structures, penetration of the onion leaf surface, and the postpenetration processes were studied using light, scanning electron, and transmission electron microscopy. Ninety-six percent of conidia germinated at 25 °C within 24 h of inoculation. Each conidium formed several germ tubes that grew in any direction across the leaf surface. Each germ tube usually terminated in a bulbous appressorium formed directly on the epidermal cell (52.4% of appressoria) or on a stoma (48.6% of appressoria). Following direct penetration of the outer epidermal cell wall or the stoma, bulbous primary hyphae developed below the appressoria. Secondary hyphae developed from the primary hyphae within 48 h after inoculation and grew within the intercellular spaces penetrating mesophyll cells. The changes in ultrastructure of cells in close proximity to hyphae and of infected cells are described. Key words: Allium cepa, electron microscopy, infection process, purple blotch.


1979 ◽  
Vol 57 (4) ◽  
pp. 324-331 ◽  
Author(s):  
R. Rohringer ◽  
W. K. Kim ◽  
D. J. Samborski

Primary leaves of wheat (Triticum aestivum L.) with and without resistance genes Sr5, Sr6, Sr8, or Sr22 were inoculated with avirulent races of stem rust (Puccinia graminis Pers. f. sp. tritici Eriks. & E. Henn.) and examined by fluorescence microscopy. In leaves containing the Sr5 gene for resistance, both epidermal and mesophyll cells fluoresced when interacting with the fungus, indicating incompatibility. In leaves containing the Sr6 gene, interacting epidermal cells did not fluoresce and incompatibility was expressed only in mesophyll cells.When the effect of the Sr5 gene was studied in leaves with different genetic backgrounds, it was found that most colonies developed only one or two haustorial mother cells in leaves containing this gene in Prelude, Marquis, or Reliance backgrounds, when examined up to 72 h after incubation. Conversely, in leaves with the Chinese Spring background, one-third of the colonies continued to grow and they produced macroscopically visible lesions. Our observations indicated that the Chinese Spring genotype partially altered the expression of the Sr5 gene in mesophyll but not in epidermal cells. In contrast, the Sr6 gene was more effective in the Chinese Spring background than in the Prelude background.Rust development in leaves with or without the Sr8 gene was the same up to 60 h after incubation, when incompatibility in resistant plants was first detected by the appearance of fluorescing host cells. By 72 h, mean colony size in resistant leaves was smaller than that in susceptible leaves, evidently because growth of runner hyphae was inhibited. Apparently, the P8 gene for avirulence was not expressed until colonies had reached considerable size. In leaves containing the Sr22 gene for resistance, the sequence of histological events was similar to that in leaves containing Sr8, but fluorescing host cells appeared later (72 h) and colony growth was inhibited only at 96 h after incubation. In both of these interactions, fluorescing host cells developed at the periphery of colonies when incompatibility was expressed. The host-parasite interaction in cells invaded before that time remained compatible even at later stages of colony development. In leaves containing the Sr5 or the Sr8 gene, but not in those with the Sr6 gene, the growth of some colonies was inhibited although they were not associated with fluorescing host cells. Evidently, host-cell necrosis was closely associated with reduced fungal growth in interactions involving Sr6, but not in interactions involving the resistance genes Sr5 and Sr8.


2006 ◽  
Vol 87 (11) ◽  
pp. 3209-3213 ◽  
Author(s):  
Sarah Cohen ◽  
Ali R. Behzad ◽  
Jeffrey B. Carroll ◽  
Nelly Panté

The parvovirus Minute virus of mice (MVM) is a small DNA virus that replicates in the nucleus of its host cells. However, very little is known about the mechanisms underlying parvovirus' nuclear import. Recently, it was found that microinjection of MVM into the cytoplasm of Xenopus oocytes causes damage to the nuclear envelope (NE), suggesting that the nuclear-import mechanism of MVM involves disruption of the NE and import through the resulting breaks. Here, fluorescence microscopy and electron microscopy were used to examine the effect of MVM on host-cell nuclear structure during infection of mouse fibroblast cells. It was found that MVM caused dramatic changes in nuclear shape and morphology, alterations of nuclear lamin immunostaining and breaks in the NE of infected cells. Thus, it seems that the unusual nuclear-import mechanism observed in Xenopus oocytes is in fact used by MVM during infection of host cells.


1979 ◽  
Vol 57 (23) ◽  
pp. 2626-2634 ◽  
Author(s):  
D. E. Harder ◽  
D. J. Samborski ◽  
R. Rohringer ◽  
S. R. Rimmer ◽  
W. K. Kim ◽  
...  

The interaction between avirulent wheat stem rust and wheat mesophyll cells containing the temperature-sensitive Sr6 gene for stem rust resistance was studied by electron microscopy. Mesophyll cells that were invaded at 26 °C (conditioning compatibility) did not develop any signs of incompatibility after they were transferred to 19 °C, at which temperature incompatibility is normally expressed. In host tissue that appeared to be invaded after the change from 26 to 19 °C, the early ultrastructural symptoms of incompatibility were a more electron-dense and often perforated invaginated host plasmalemma, disruptions of the noninvaginated host plasmalemma, vacuolation of the cytoplasm, and accumulations of electron-dense material along the membranes of the vacuoles. At later stages in the development of incompatible interactions, the electron-dense accumulations along the vacuole membranes increased in size and occurred along chloroplast and mitochondrial membranes. Eventually, the entire protoplasts were electron dense and collapsed. In haustoria and haustorial mother cells, incompatibility was usually expressed by a uniform increase in electron density of the cytoplasm. In the Sr6/P6 interaction at 19 °C, host cell necrosis was not always accompanied by fungal necrosis or vice versa. In Sr5/P5 interactions, which were examined for comparison, the intracellular symptoms of incompatibility were similar to those of the Sr6/P6 interactions.


1975 ◽  
Vol 21 (12) ◽  
pp. 1930-1939 ◽  
Author(s):  
Darrell A. Kinden ◽  
Merton F. Brown

Scanning- and transmission-electron microscopy were used to examine developing and mature functional arbuscules in mycorrhizal roots of yellow poplar. Arbuscules developed from intracellular hyphae which branched repeatedly upon penetration into the host cells. Intermediate and late stages of development were characterized by the production of numerous, short, bifurcate hyphae throughout the arbuscule. Mature arbuscules exhibited a coralloid morphology which resulted in a considerable increase in the surface area of the endophyte exposed within the host cells. Distinctive ultrastructural features of arbuscular hyphae included osmiophilic walls, nuclei, abundant cytoplasm, glycogen, and numerous small vacuoles. All arbuscular components were enclosed by host wall material and cytoplasm during development and at maturity. In infected cells, host nuclei were enlarged and the cytoplasm associated with the arbuscular branches typically contained abundant mitochondria, endoplasmic reticulum, and proplastids. Ultrastructural observations suggested that nutrient transfer may be predominantly directed toward the fungal endophyte during arbuscular development and while mature arbuscules remain functional.


1974 ◽  
Vol 52 (5) ◽  
pp. 1107-1115 ◽  
Author(s):  
R. A. Skipp ◽  
D. J. Samborski

Seedling leaves of resistant (Sr6) and susceptible (sr6) near-isogenic wheat lines inoculated with urediospores of Puccinia graminis f. sp. tritici race C 17 (56) became infected at similar rates. Host cells of the resistant line became necrotic after haustorial penetration (beginning about 20 h after inoculation), whereas necrosis was rarely seen in the susceptible line and colonies grew to form sporulating pustules. Some colonies in the resistant line appeared to have stopped growing by about 60 h after inoculation, while others grew slowly, the area of necrosis increasing as they expanded.Inoculated resistant-line plants became susceptible when incubated at 25 °C rather than 20 °C. Provided that the plants were kept at 25 °C for at least 1 day before inoculation, no host cell became necrotic. The necrotic response was resumed, and a more resistant infection type developed when infected seedlings were transferred from 25 °C to 20 °C. The converse occurred when resistant plants were grown and incubated at 20 °C, then transferred to 25 °C.Effects on fungal growth and the action of the Sr6 gene were considered to be closely associated with host cell necrosis. Temperature sensitivity appeared to be a property of the host plant.


1977 ◽  
Vol 23 (7) ◽  
pp. 873-883 ◽  
Author(s):  
B. Bassett ◽  
R. N. Goodman ◽  
A. Novacky

Ineffective nodules, formed on Clark-63 soybeans by Rhizobium japonicum strain 8-0 (Iowa), and effective nodules, formed on Clark-63 soybeans by strain 138 (U.S.D.A.), have been examined by electron microscopy at 10–12, 16, and 21 days after seed inoculation. Though strain 8-0 bacteria are able to infect the host cells in a normal manner, infection is followed closely by a progressive deterioration of the symbiosis involving selective autolysis of host cell contents and degeneration of the intracellular bacteria. The host cells, though disrupted, apparently survive the destruction of the bacteria. The observed ultrastructural changes suggest either a suddenly manifested incompatibility between host and rhizobia, or an acute, localized, nitrogen starvation in the infected cells.


1986 ◽  
Vol 64 (3) ◽  
pp. 626-631 ◽  
Author(s):  
H. D. M. Gousseau ◽  
B. J. Deverall

The development of avirulent and virulent strains of stem rust (Puccinia graminis Pers. f.sp. tritici Eriks. & Henn.) in a susceptible wheat line and two cultivars bearing the Sr15 allele for resistance was studied, mainly by fluorescence microscopy. Formation of appressoria, substomatal vesicles, infection hyphae, and the first haustorium was unaffected by resistance. The first effect of Sr15 expression was a slower rate of haustorial mother cell formation and was first seen 48 h after inoculation. Effects on hyphal branching and colony radii followed. Necrosis of host cells was first seen at 42 h, but inspection of individual infection sites showed that necrosis did not coincide with effects on haustorial mother cells. It is possible that deterioration of host cells leading to visible host cell necrosis may be related to effects on rust development. Sr15 expression gave a mesothetic reaction, first seen microscopically 60 h after inoculation. Differences between individual infection sites in this reaction may be related to the timing of the onset of necrosis.


Author(s):  
R. Stephens ◽  
K. Traul ◽  
D. Woolf ◽  
P. Gaudreau

A number of antigens have been found associated with persistent EBV infections of lymphoblastoid cells. Identification and localization of these antigens were principally by immunofluorescence (IF) techniques using sera from patients with nasopharyngeal carcinoma (NPC), Burkitt lymphoma (BL), and infectious mononucleosis (IM). Our study was mainly with three of the EBV related antigens, a) virus capsid antigen (VCA), b) membrane antigen (MA), and c) early antigens (EA) using immunoperoxidase (IP) techniques with electron microscopy (EM) to elucidate the sites of reactivity with EBV and EBV infected cells.Prior to labeling with horseradish peroxidase (HRP), sera from NPC, IM, and BL cases were characterized for various reactivities by the indirect IF technique. Modifications of the direct IP procedure described by Shabo and the indirect IP procedure of Leduc were made to enhance penetration of the cells and preservation of antigen reactivity.


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