Specific polyclonal antibodies for the obligate plant parasite Polymyxa - a targeted recombinant DNA approach

A - Papers appearing in refereed journals

Mutasa-Göttgens, E. S., Chwarszczynska, D. M., Halsey, K. and Asher, M. J. C. 2000. Specific polyclonal antibodies for the obligate plant parasite Polymyxa - a targeted recombinant DNA approach. Plant Pathology. 49 (2), pp. 276-287. https://doi.org/10.1046/j.1365-3059.2000.00446.x

AuthorsMutasa-Göttgens, E. S., Chwarszczynska, D. M., Halsey, K. and Asher, M. J. C.
Abstract

Highly specific rabbit polyclonal antibodies for the obligate sugar-beet root parasite, Polymyxa betae, were produced using a novel recombinant DNA approach. Parasite cDNA was selectively isolated from infected roots, expressed in vitro, and the purified protein used to raise antibodies. This produced clean, precisely targeted antibodies, and allowed for rigorous screening of candidate genes and their products at the molecular level prior to animal immunization. This approach selects for genes whose products are highly expressed by the parasite in planta, and five such candidate genes from Polymyxa betae were identified and cloned. Polyclonal antiserum developed using the product of one such gene was found to react specifically with P. betae in sugar-beet roots and with the closely related Polymyxa graminis in barley roots, and to cross-react with Plasmodiophora brassicae in cabbage roots, without the need for further purification. No cross-reaction was detected with protein extracts from potato roots infected by the plasmodiophoromycete Spongospora subterranea. In all cases, there was no interaction with proteins from host plants, or from other microorganisms found in association with uninoculated sugar-beet, barley, cabbage and potato roots.

KeywordsAgronomy; Plant Sciences
Year of Publication2000
JournalPlant Pathology
Journal citation49 (2), pp. 276-287
Digital Object Identifier (DOI)https://doi.org/10.1046/j.1365-3059.2000.00446.x
Open accessPublished as non-open access
FunderBiotechnology and Biological Sciences Research Council
Funder project or code451
Project: 404091
PublisherWiley
ISSN0032-0862

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