PRODUCTION AND USE OF MONOCLONAL-ANTIBODIES TO PSEUDOMONAS-ANDROPOGONIS

Pseudomonas andropogonis is an important pathogen of worldwide distribution in ornamental and other plant species from 15 families. This paper reports the production and characterization of monoclonal antibodies (MAbs) to P. andropogonis and evaluation of their use in the detection of the pathogen in carnation cuttings. Ten stable hybridoma cell lines were produced. Results of indirect ELISA and indirect immuno-fluorescence showed that MAb 6B3 was specific for P. andropogonis; MAb 3D5W1 reacted with both P. andropogonis and P. caryophylli; six other MAbs reacted with all strains of seven species of rRNA group II pseudomonads tested except P. solanacearum and P. pickettii. Eight of the ten MAbs failed to cross-react with other non-fluorescent or fluorescent pseudomonads, xanthomonads and other bacteria tested. P. andropogonis was similar in protein profile to other rRNA group II pseudomonads tested except P. solanacearum and P. pickettii. Epitopes were clearly located within the cell by immunogold labelling. Of four MAbs that were isotyped, two possessed IgG1 and two the IgM heavy chain. P. andropogonis was readily detected by combining immunofluorescence and detached carnation leaf assay using an initial inoculum of 4 x 100 colony forming units (cfu) ml-1 after enrichment at room temperature for 4 days.