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  • 1
    Electronic Resource
    Electronic Resource
    Caractérisation directe deRhizoctonia solani Kühn AG3 à partir de sclérotes sur tubercules de pomme de terre par deux méthodes sérologiques (1993)
    Springer
    Potato research 36 (1993), S. 43-53 
    Details
    ISSN: 1871-4528
    Keywords: chromatographie ; électrophorèse ; immunodiffusion ; ELISA indirect ; groupes d'anastomoses ; immunisation rapide
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Description / Table of Contents: Summary Total mycelial extract ofR. solani AG3 was purified by ion-exchange chromatography. Beforehand part of the extract was heat-treated at 60°C for 10 min (Ch) and the rest left untreated (N). Results of the chromatographic analysis are given in Fig. 3. Electrophoretic analysis of the peak fractions is shown in Fig. 2. Peak C was chosen as antigen for the preparation of the two antisera by intravenous injections in two rabbits. Antibodies obtained were titrated in both cases by immunodiffusion and in indirect ELISA (Fig. 4). Specificity was examined by immunodiffusion and ELISA on heat-treated extracts of reference strains listed in Table 1. Similar results were obtained for the two antisera which were both specific to AG3 (Fig. 1 and Table 2). Characterisation tests were done by the two methods directly on sclerotia of tubers of different cultivars and origins harvested in 1991 (Table 3). The purified antigens appeared to be of a complex nature (peptides-polysaccharides) and were very immunogenic. Antibodies raised against them were sufficiently specific as to allow the identification ofR. solani AG3 sclerotica on tubers.
    Notes: Résumé Les antisérums sont obtenus en immunisant deux lapins par voie intraveineuse avec des antigènes purifiés par chromatographie d'échange d'ions. On utilise deux préparations d'extraits mycéliens: normale et chauffée. La spécificité des anticorps obtenus est appréciée sur différentes souches deRhizoctonia. Deux techniques sérologiques sont utilisées: immunodiffusion et ELISA indirect. Les deux immunsérums obtenus présentent la même spécificité vis-à-vis du groupe AG3. Les deux techniques permettent la reconnaissance du groupe AG3 à partir des sclérotes formés sur tubercules.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF02359833
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  • 2
    Electronic Resource
    Electronic Resource
    Utilisation d'anticorps spécifiques d'une fraction obtenue par chromatographie d'exclusion pour une amélioration de la caractérisation sérologique deRhizoctonia solani Kühn AG3 (1991)
    Springer
    Potato research 34 (1991), S. 149-158 
    Details
    ISSN: 1871-4528
    Keywords: immunodiffusion ; électrotransfert ; marquage immunoenzymatique ; groupes d'anastomose ; Solanum tuberosum L.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Description / Table of Contents: Summary A soluble mycelial extract ofR. solani AG3 was purified by size exclusion chromatography. Five peaks were present in the elution profile: the first was well defined and of large volume. It was retained to immunise a female rabbit by intradermic injection. The titre of the antiserum determined by double immuno-diffusion in agarose was 0.125. The antiserum was tested on isolates ofR. solani AG1, AG2, AG3, AG4, AG5, AG6, AG7, AGB1 and ofCeratobasidium CAG1, CAG2, CAG3, CAG4 and CAG5 (Table 1) using three different methods: - immunodiffusion with the test material adjusted to 3 mg/ml of proteins. Only AG3 isolates formed a single precipitation line (Fig. 1). - electrotransfer and immuno-blotting. Test samples containing 8 mg/ml of proteins were electrophoresed under natural conditions in a discontinuous system. The proteins were then electrotransferred to a nitrocellulose membrane. Immunoenzymatic staining showed that only one band was present in AG3 isolates (Fig. 2). - immunoenzymatic labelling of the mycelium demonstrated the presence of dark granules on the cell wall of only AG3 isolates (Fig. 3).
    Notes: Résumé La purification d'un extrait mycélien soluble d'une souche deR. solani AG3 par chromatographie d'exclusion permet de recueillir une fraction antigénique. Une lapine est immunisée par voie intradermique. L'immunsérum titre 0,125 par la technique de double-diffusion en agarose. Différentes techniques sérologiques (immunodiffusion, électrotransfert suivi de révélation des fractions antigéniques, coloration immunoenzymatique du mycélium) sont appliquées à des souches deR. solani AG1, AG2, AG3, AG4, AG5, AG6, AG7, AGBI et deCeratobasidium CAG1, CAG2, CAG3, CAG4 et CAG5. Le sérum se révèle spécifique des souches AG3.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF02358036
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