Antibodies from Chicken Eggs as Probes for Antigens from Pasteuria penetrans Endospores

S. Y. Chen, J. Charnecki, J. F. Preston, D. W. Dickson, J. D. Rice

Abstract


The bacteria Pasteuria spp. have been identified as among the most promising of several microbial organisms currently under investigation as biological control agents of plant-parasitic nematodes. As part of our goal to develop methods to discriminate isolates of Pasteuria penetrans with different host preferences, we investigated the potential of developing antibody probes to identify endospores of different isolates of P. penetrans. Polyclonal IgY antibodies were raised in chickens against endospores of P. penetrans isolates P20 and P100. Hens were injected with P20 or P100 endospore suspensions and boosted at 14 days. Anti-spore titers were determined with ELISA on yolk extracts of individual eggs as a function of time. The highest titers were found in eggs produced at 22 to 35 days after initial injections. Yolk extracts showing the highest titers were combined and processed to provide partially purified IgY preparations. SDS-PAGE and immunoblot analyses identified protein antigens with M[subr] values of 23-24, 46, and 57-59 KDa common to both P20 and P100 endospores. One protein antigen with an M[subr] value of 62 KDa was unique to the PI00 endospores. The IgY antibodies reduced the attachment of Pastcuria endospores to their nematode hosts, indicating antibody interaction with antigens on the endospore surface that are involved in the recognition and attachment processes. Key words: antibody, antigen, bacterium, biological control, ELISA, endospore, immunoblot, Meloidogyne arenaria, Meloidogyne incogwita, Meloidogyne javanica, Pasteuria penetrans, protein, root-knot nematode, SDS-PAGE.

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