Development of a recombinant antibody ELISA test for the detection ofPolymyxa betaeand its use in resistance screening

Abstract

An ELISA test was developed for the quantitative detection of the obligate parasite Polymyxa betae , the vector of Beet necrotic yellow vein virus (BNYVV), in infected sugarbeet roots. The test used monoclonal and polyclonal antibodies raised to a recombinantly expressed glutathione-S-transferase (GST) from P. betae . A close correlation was found between the number of P. betae zoospores in serially diluted suspensions and absorbance values in the ELISA test. Timecourse studies of plants grown in naturally infested soils in controlled environment tests demonstrated the value of the ELISA test in screening for P. betae resistance. In preliminary tests, P. betae -resistant accessions of the wild sea beet ( Beta vulgaris ssp. maritima ), which might be used to restrict the transmission of BNYVV, were identified.

Management of plant health risks associated with processing of plant-based wastes: A review

“…A rapid method for the detection of Polymyxa DNA directly from soil reported by Ward et al (2004) was capable of detecting as few as 2.78 × 10 3 cysts of P. betae g -1 soil, but was unable to distinguish viable from nonviable cystosori. Kingsnorth et al (2003) developed a recombinant antibody ELISA assay for the P. betae glutathione-S-transferase, which is expressed during infection.…”

“…Phytophthora kernoviae sporangia oospores similar to P. ramorum no data Schena et al (2006) Polymyxa spp cystosori in roots or soil 100-150 cystosori detection of spores from soil using Real Time PCR 1.39 × 10 3 cystosori g -1 soil Ward et al (2004) bait tests followed by glutathione-S-transferase ELISA or microscopy no data √ Kingsnorth et al (2003) MPN method bait test method 50-100 cystosori √ Tuitert (1990) 58 Roenhorst et al (2005) …”

Detection and molecular characterization of Polymyxa betae, transmitting agent of sugar beet rhizomania disease, in Iran

“…ELISA is ideal for usage in P. betae screening in sugar beet germplasm. Unlike PCR-based methods, antibodies detect the actual amount of antigen without amplification, and in addition are more likely able to differentiate between live and dead pathogen biomass (Kingsnorth et al, 2003). ELISA sensitivity, as determined by an ability to detect low levels of P. betae infection, enhances their utility for screening f ield samples.…”

“…To detect P. betae and BNYVV, DAS-ELISA test was carried out as described by Kingsnorth et al (2003) and Asher et al (2002) respectively, with slight modifications. Lateral root samples collected from Regina plants were used for P. betae detection and BNYVV quantification.…”

“…Previous study showed that GST was expressed at high levels in P. betae, and could overcome host plant defense mechanisms (Mutasa-Gottgens et al, 2000). Kingsnorth et al (2003) found that P. betae GST is an immunogenic protein. Safarpour et al (2012) cloned the coding sequences of the GST gene from an Iranian P. betae isolate and developed an efficient procedure for producing a polyclonal antibody against this Iranian isolate.…”