DEVELOPMENT OF A POLYCLONAL ANTIBODY-BASED IMMUNOASSAY FOR THE EARLY DETECTION OF SCLEROTINIA-SCLEROTIORUM IN RAPESEED PETALS

A serological test has been developed that allows the early detection of infection of young petals by Sclerotinia sclerotiorum, an important pathogen of rapeseed. Two steps were required to obtain an antiserum sufficiently specific for S. sclerotiorum. Soluble mycelial extracts of S. sclerotiorum were used to produce the first generation polyclonal antiserum. This was not specific for S. sclerotiorum in double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) and allowed the screening of cross-reacting fungal species such as Botrytis cinerea, a pathogen commonly present on rapeseed petals. Using a polyclonal anti-B. cinerea serum enabled the absorption, by serial cycles, of S. sclerotiorum antigens common to B. cinerea. Residual antigens were then used as immunogens for the production of two second generation antisera (S1 and S2) which were then tested by DAS-ELISA. Cross-reactions with B. cinerea decreased with purification cycles of the immunogen whereas cross-reactions with some unrelated fungi slightly increased. S. sclerotiorum and B. cinerea were distinguishable using antiserum S2.