Analysis of differentiation and development of the specialized infection structures formed by biotrophic fungal plant pathogens using monoclonal antibodies

1995 ◽  
Vol 73 (S1) ◽  
pp. 408-417 ◽  
Author(s):  
Jonathan R. Green ◽  
Naomi A. Pain ◽  
Martin E. Cannell ◽  
Calum P. Leckie ◽  
Sharon McCready ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Plasma Membrane ◽  
Powdery Mildew ◽  
Plant Pathogens ◽  
Colletotrichum Lindemuthianum ◽  
Cdna Expression Library ◽  
Erysiphe Pisi ◽  
Infection Structures ◽  
Fungal Plant Pathogens

Monoclonal antibodies have been used to study the differentiation and development of the specialized infection structures formed in the Colletotrichum–bean and powdery mildew – pea interactions. In the Colletotrichum lindemuthianum – bean interaction, monoclonal antibodies have been used to show that the extracellular matrices associated with conidia, germ tubes, and appressoria differ in composition and that the extracellular glycoproteins are organized into specific regions of the fungal cell surface. Monoclonal antibody UB27 has been used to show that the plasma membrane of appressoria is differentiated into distinct domains, with the integral membrane glycoprotein identified by UB27 being excluded from the pore region. UB25 recognizes a glycoprotein located specifically in the cell wall/matrix of intracellular hyphae and is expressed only during the biotrophic phase of development. In the Erysiphe pisi – pea interaction, UB8 and UB10 identify glycoproteins specific to the haustorial plasma membrane within the haustorial complex. Monoclonal antibodies that recognize the extrahaustorial membrane have shown that this membrane contains specific components, as well as glycoproteins in common with the host plasma membrane. UB8 has been successfully used to isolate a gene sequence coding for the protein antigen, by immunoscreening a cDNA expression library prepared from infected epidermis. An antibody that recognizes the plant endoplasmic reticulum has been used to show that this structure reorganizes around the developing haustorial complex in pea epidermal cells. Key words: appressorium, biotrophy, Colletotrichum lindemuthianum, Erysiphe pisi, haustorium, monoclonal antibody, powdery mildew.

scholarly journals Isolation and characterization of monoclonal antibodies directed against plant plasma membrane and cell wall epitopes: identification of a monoclonal antibody that recognizes extensin and analysis of the process of epitope biosynthesis in plant tissues and cell cultures.

1988 ◽  
Vol 107 (1) ◽  
pp. 163-175 ◽  
Author(s):  
D J Meyer ◽  
C L Afonso ◽  
D W Galbraith
Keyword(s):  
Monoclonal Antibody ◽  
Cell Wall ◽  
Monoclonal Antibodies ◽  
Plasma Membrane ◽  
Solid Phase ◽  
Culture Media ◽  
Gradient Centrifugation ◽  
Cell Suspension Cultures ◽  
Isolation And Characterization

Membranes from tobacco cell suspension cultures were used as antigens for the preparation of monoclonal antibodies. Use of solid phase and indirect immunofluorescence assays led to the identification of hybridomas producing antibodies directed against cell surface epitopes. One of these monoclonal antibodies (11.D2) was found to recognize a molecular species which on two-dimensional analysis (using nonequilibrium pH-gradient electrophoresis and SDS-PAGE) was found to have a high and polydisperse molecular mass and a very basic isoelectric point. This component was conspicuously labeled by [3H]proline in vivo. The monoclonal antibody cross-reacted with authentic tomato extensin, but not with potato lectin nor larch arabinogalactan. Use of the monoclonal antibody as an immunoaffinity reagent allowed the purification of a tobacco glycoprotein which was identical in amino acid composition to extensin. Finally, immunocytological analyses revealed tissue-specific patterns of labeling by the monoclonal antibody that were identical to those observed with a polyclonal antibody raised against purified extensin. We have concluded that monoclonal antibody 11.D2 recognizes an epitope that is carried exclusively by extensin. Analysis of cellular homogenates through differential and isopycnic gradient centrifugation revealed that biosynthesis of the extensin epitope was found on or within the membranes of the endoplasmic reticulum, Golgi region and plasma membrane. This result is consistent with the progressive glycosylation of the newly-synthesized extensin polypeptide during its passage through a typical eukaryotic endomembrane pathway of secretion. The 11.D2 epitope was not found in protoplasts freshly isolated from leaf tissues. However, on incubation of these protoplasts in appropriate culture media, biosynthesis of the epitope was initiated. This process was not impeded by the presence of chemicals that are reported to be inhibitors of cell wall production or of proline hydroxylation.

Topographical signaling for cell differentiation in the plant pathogenic fungus, Uromyces

1989 ◽  
Vol 47 ◽  
pp. 784-785
Author(s):  
H.C. Hoch
Keyword(s):  
Plant Pathogens ◽  
Developmental Process ◽  
Pathogenic Fungus ◽  
Host Cells ◽  
Subsequent Infection ◽  
Infection Structures ◽  
Rust Diseases ◽  
Fungal Plant Pathogens ◽  
Plant Signals ◽  
Important Structure

Fungal plant pathogens invade host cells with a variety of specialized infection structures, however, for most fungi the appressorium is developmentally the first and most important structure to be formed in preparation for host colonization. It must be positioned at an appropriate site on the host in a timely way so that subsequent infection can be assured. For fungi which cause rust diseases of plants, positioning the appressorium is the most critical stage because invasion of the host can occur only via the stomata. Uredospores of these fungi (e.g.,Uromyces appendiculatus) germinate and grow, directed by the leaf (bean) surface topography toward stomata where they cease growth and develop appressoria directly over the stomatal openings. Development of the appressorium is accompanied by ameboid-like migration of the cytoplasm into the ballooning hyphal tip, DNA synthesis and nuclear division, synthesis of several “differentiation” proteins, and a rearrangement of the cytoskeleton. An orderly succession of subsequent infection structures (e.g., infection pegs, vesicles) follow in a preprogrammed sequence once the initial developmental process has been started.My research goals have been to determine what feature(s) of the host plant signals infection structure formation and how the fungus perceives these signals.

scholarly journals Monoclonal antibodies that immunoreact with a cation-stimulated plant membrane ATPase

1982 ◽  
Vol 203 (1) ◽  
pp. 51-54 ◽  
Author(s):  
J J C Chin
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Plasma Membrane ◽  
Marker Enzyme ◽  
Membrane Atpase ◽  
Hybridoma Technology ◽  
Membrane Antigens ◽  
Monospecific Antibodies

Hybridoma technology has been used successfully to generate monoclonal-antibody probes against protoplast membrane antigens. Hybridomas secreting monoclonal antibodies that either inhibit or stimulate a putative plasma-membrane marker enzyme, (K+ + Mg2+)-stimulated pH 6.5 ATPase, have been identified and cloned. The specificity of monoclonal-antibody probes on the activity of other phosphate-hydrolysing enzymes has also been examined. The production and identification of monospecific antibodies capable of immunoreacting with particular component proteins in a complex plant membrane mixture highlight the usefulness of hybridoma methodology for the enzymologist, especially since such monoclonal antibodies can be used in the purification of proteins by immunoaffinity techniques.

Rapid quantification and in situ detection of nitrifying bacteria in biofilms by monoclonal antibody method

2000 ◽  
Vol 41 (4-5) ◽  
pp. 301-308 ◽  
Author(s):  
N. Noda ◽  
H. Ikuta ◽  
Y. Ebie ◽  
A. Hirata ◽  
S. Tsuneda ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Fluorescent Antibody ◽  
Nitrifying Bacteria ◽  
Fluorescent Antibody Technique ◽  
Antibody Technique ◽  
Antibody Method ◽  
In Situ Detection ◽  
Rapid Quantification

Fluorescent antibody technique by the monoclonal antibody method is very useful and helpful for the rapid quantification and in situ detection of the specific bacteria like nitrifiers in a mixed baxterial habitat such as a biofilm. In this study, twelve monoclonal antibodies against Nitrosomonas europaea (IFO14298) and sixteen against Nitrobacter winogradskyi (IFO14297) were raised from splenocytes of mice (BALB/c). It was found that these antibodies exhibited little cross reactivity against various kinds of heterotrophic bacteria. The direct cell count method using monoclonal antibodies could exactly detect and rapidly quantify N. europaea and N. winogradskyi. Moreover, the distribution of N. europaea and N. winogradskyi in a biofilm could be examined by in situ fluorescent antibody technique. It was shown that most of N. winogradskyi existed near the surface part and most of N. europaea existed at the inner part of the polyethylene glycol (PEG) gel pellet, which had entrapped activated sludge and used in a landfill leachate treatment reactor. It was suggested that this monoclonal antibody method was utilized for estimating and controlling the population of nitrifying bacteria as a quick and favorable tool.

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