Utilisation d'anticorps spécifiques d'une fraction obtenue par chromatographie d'exclusion pour une amélioration de la caractérisation sérologique deRhizoctonia solani Kühn AG3

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.

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).