Abstract
Erwinia stewartii (Dye) causes Stewart's Wilt, a seed transmitted bacterial disease of maize. This pathogen, of quarantine importance, is responsible for serious crop losses in many tropical countries. At present, seed health tests are time consuming and lack sensitivity. This paper describes the development of a polymerase chain reaction (PCR) test to identify E. stewartii and the comparison of this with two other seed health testing methods (nigrosine selective medium and ELISA). PCR with arbitrary primers was used to identify a DNA fragment for use as a probe to detect E. stewartii isolated from maize seed. Specific nested primers for E. stewartii were designed from the partial sequence of the E. stewartii probe. These primers were highly specific and did not cross-react with other Erwinias or a wide of range of bacterial isolates obtained from maize seed samples. Identification of this pathogen from cultured cells is rapid using PCR and can be done in less than a working day (5 hours). The sensitivity of the nested PCR is sufficient to amplify from approximately ten bacterial cells.
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Claims 4
Claim · model-written summary
The primers were highly specific for E. stewartii.
Zea maysNot reportedInteractor
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“These primers were highly specific and did not cross-react with other Erwinias or a wide of range of bacterial isolates obtained from maize seed samples”
Claim · model-written summary
The primers did not cross react with other Erwinias or bacterial isolates from maize seeds.
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“did not cross-react with other Erwinias or a wide of range of bacterial isolates obtained from maize seed samples”
Claim · model-written summary
PCR identification from cultured cells took less than five hours.
Not reportedTaxon
Verbatim
e0“Identification of this pathogen from cultured cells is rapid using PCR and can be done in less than a working day (5 hours)”
Claim · model-written summary
Nested PCR amplified from approximately ten bacterial cells.
Not reportedTaxon
Verbatim
e0“The sensitivity of the nested PCR is sufficient to amplify from approximately ten bacterial cells”