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  • 1
    Electronic Resource
    Electronic Resource
    Development of monoclonal antibody-based immunological assays for the detection of live propagules of Rhizoctonia solani in soil (1993)
    Oxford, UK : Blackwell Publishing Ltd
    Plant pathology 42 (1993), S. 0 
    Details
    ISSN: 1365-3059
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Mouse monoclonal antibodies (mAbs) and rabbit (polyclonal) antiserum were used to develop DIAGNOSTIC-ELISA, double-antibody-sandwich-ELISA (DAS-ELISA), DIP-STICK and immuno-fluorescence colony staining immunoassays for the specific detection of Rhizoctonia solani in soil. mAbs were raised against an anastomosis group 4 isolate of R. solani. Mice were immunized using either phosphate-buffered saline (PBS) suspensions of lyophilized mycelium plus Quil A adjuvant, or with a solubilized acetone precipitate prepared from cell-free surface washings from solid slant cultures. Polyclonal antisera were raised in rabbits using PBS suspensions of lyophilized mycelium and Quil A adjuvant. Hybridoma supernatants and rabbit antisera were screened by ELISA. Four of the cell lines raised produced mAbs that were species-specific. They recognized antigens from R. solani by ELISA and immunofluorescence, but not other related or unrelated species of soil-borne fungi. The remaining cell line produced mAbs that cross-reacted slightly, by ELISA, with antigens from R. cerealis. These mAbs did not recognize R. cerealis by immunofluorescence, or other related or unrelated soil-borne fungi, by ELISA.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1365-3059.1993.tb01563.x
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  • 2
    Electronic Resource
    Electronic Resource
    Comparison of isoenzyme and DNA markers for differentiating W-, R- and C-pathotypes of Pseudocercosporella herpotrichoides (1992)
    Oxford, UK : Blackwell Publishing Ltd
    Plant pathology 41 (1992), S. 0 
    Details
    ISSN: 1365-3059
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Biochemical differences between the wheat, rye and couch-grass pathotypes of the eyespot pathogen, Pseudocercosporella herpotrichoides, have been identified by comparing isoenzyme profiles and DNA markers from several isolates. The wheat (W) and rye (R) pathotypes were clearly differentiated by both techniques, with isoenzyme profiling also separating the couch-grass isolates from the W- and R-type isolates. The isoenzyme profiles separated the P. herpotrichoides pathotypes from the two related species, P. anguioides and P. aestiva. The isoenzyme profiles of P. anguioides were closer to those of the R-type isolates of P. herpotrichoides than the W-type isolates, whereas the banding pattern of P. aestiva was very different. The isoenzyme profiles of the two German isolates, originally obtained from Nirenberg's laboratory, P. herpotrichoides var. herpotrichoides and var. acuformis, were similar to those of the wheat and rye pathotypes, respectively, isolated from UK-infected material.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1365-3059.1992.tb02458.x
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  • 3
    Electronic Resource
    Electronic Resource
    Development of a monoclonal antibody immunoassay for the eyespot pathogen Pseudocercosporella herpotrichoides (1993)
    Oxford, UK : Blackwell Publishing Ltd
    Plant pathology 42 (1993), S. 0 
    Details
    ISSN: 1365-3059
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: A double-antibody-sandwich ELISA test has been developed for the detection of Pseudocercosporella herpotrichoides using a highly specific monoclonal antibody PH-10 as the capture antibody and genus-specific rabbit polyclonal antiserum as the detector antibody. The assay recognizes extracts from plants both artificially and naturally infected with P. herpotrichoides giving at least three-fold higher absorbance values with extracts from Pseudocercosporella-infected tissue than with extracts from healthy tissues or from tissues naturally infected with Microdochium nivale, Rhizoctonia cerealis or material artificially inoculated with P. anguioides. The assay tested positively against all isolates of P. herpotrichoides, including both W-type and R-type isolates. In this assay system, extraction of the antigen from the stem bases of infected plants is a one-step process not requiring any dilution procedures. The assay can be used to detect the pathogen in presymptomatic infected seedlings. The immunogen used to generate the specific monoclonal antibody and the rabbit antiserum was a mycelial extract from which the high-molecular-weight proteins and glycoproteins had been removed by ammonium sulphate precipitation. The high-molecular-weight fraction was shown to contain cross-reactive antigens; it induced antiserum in mice that cross-reacted with the other stem-base fungi even at high dilutions. The monoclonal antibody PH-10 is an IgM antibody. Heat and periodate treatment of the antigen indicate that it is a glycoprotein and that the epitope recognized by the antibody is a protein.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1365-3059.1993.tb01518.x
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  • 4
    Electronic Resource
    Electronic Resource
    Development of a monoclonal antibody-based immunodetection assay for Botrytis cinerea (1992)
    Oxford, UK : Blackwell Publishing Ltd
    Plant pathology 41 (1992), S. 0 
    Details
    ISSN: 1365-3059
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Three hybridoma cell lines secreting antibodies that specifically recognize Botrytis cinerea and B.fabae, but not B. allii, have been raised from splenocytes of mice immunized with a low molecular-weight fraction (30 kDa) from surface washings of B. cinerea. Antibodies from these cell lines have been used to develop an antigen-based elisa test that will detect B. cinerea in strawberries. This monoclonal antibody immunoassay detection assay should prove useful to both the cut-flower and wine industries. Supernatants from the three specific cell lines recognize mycelial fragments, saline extracts of mycelia and germinating conidia by both ELISA and immunofluorescence. Recognition of non-germinating spores is poor. Supernatants from the specific cell lines did not recognize other fungi normally involved in post-harvest spoilage of fruits and vegetables. Supernatants from KH4 gave the lowest background values with healthy tissue. Indirect evidence from heat, protease and periodate treatment of the antigens indicates that antibodies from all three specific cell lines are recognizing carbohydrate epitopes on a glycoprotein.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1365-3059.1992.tb02442.x
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  • 5
    Electronic Resource
    Electronic Resource
    Monoclonal antibodies to specific components of the Dutch elm disease pathogen Ophiostoma ulmi (1989)
    Oxford, UK : Blackwell Publishing Ltd
    Plant pathology 38 (1989), S. 0 
    Details
    ISSN: 1365-3059
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: The potential of polyclonal antisera and monoclonal antibodies to differentiate the EAN and NAN aggressive subgroups of Ophiostoma ulmi was explored. Polyclonal antisera, when tested by ELISA, cross-reacted widely with unrelated species and failed to distinguish between the two aggressive subgroups but small quantitative differences were found, particularly between antigens secreted overnight, by EAN and NAN germlings. Monoclonal antibodies were raised in mice against mycelial homogenates. From two fusions, 33 cell lines were raised that secreted antibodies positive for O. ulmi. Approximately one third were non-specific; 11 were specific either to species or subspecies. Two cell lines differentiated mycelial antigens of the aggressive isolates of O. ulmi from those of the non-aggressive subgroup, but not antigens from surface washings. Only quantitative differences were detected between the EAN and NAN aggressive subgroups. Almost all the monoclonal antibodies and antiserum recognized antigens present in surface washings of cultures on solid medium, in cell-free extracts of mycelial homogenates, in cell-free culture fluids, and in substances secreted overnight by germinating spores. Specific detection of such molecules promises to provide a highly sensitive mechanism for studying early pathogen/host plant interactions. Most of the monoclonal antibodies appeared to have potential diagnostic value; they gave readings twofold to tenfold higher with extracts from diseased than from healthy tissue. However, one cell line that secreted antibodies specific to O. ulmi cross-reacted strongly with extracts of healthy tissue.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1365-3059.1989.tb01422.x
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  • 6
    Electronic Resource
    Electronic Resource
    Development of ELISA for Ophiostoma ulmi using antigen-coated wells (1988)
    Oxford, UK : Blackwell Publishing Ltd
    Plant pathology 37 (1988), S. 0 
    Details
    ISSN: 1365-3059
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: A simple sensitive ELISA test has been developed that detects either antibodies to, or antigens of, the Dutch elm pathogen, Opiostoma (Ceratocystis) ulmi. The dilution end point for antisera raised in mice againt mycelial antigens was 10-4. The minimum amount of antigen detected in wells passively coated with antigens solubilized in phosphate-buffered saline was 500 pg/ml. The assay proved an effective method for rapidly screening large numbers of hybridoma supernatants for monoclonal antibodies that recognize O. ulmi antigens and it has also been used to detect fungal antigens in saline extracts of diseased tissue. Non-specific binding of preimmune antiserum to plant extracts was markedly higher in extracts from diseased than non-diseased tissue. Production by the diseased host or by the pathogen in vivo of a protein A or lectin-like molecule that binds non-specifically to immunoglobulins is postulated.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1365-3059.1988.tb02192.x
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  • 7
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    The intersection between cell wall disassembly, ripening, and fruit susceptibility to Botrytis cinerea (2008)
    National Academy of Sciences
    Details
    Publication Date: 2008-01-16
    Print ISSN: 0027-8424
    Electronic ISSN: 1091-6490
    Topics: Biology , Medicine , Natural Sciences in General
    Published by National Academy of Sciences
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