Identification and in planta detection of Pseudomonas syringae pv. tomato using PCR amplification of hrpZPst |
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| Authors: | Massimo Zaccardelli Annalisa Spasiano Carlo Bazzi and Massimo Merighi |
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| Institution: | (1) Istituto Sperimentale per le Colture Industriali, Mi.P.A.F., SS 18 no 156, Battipaglia (SA), 84091, Italy;(2) Dipartimento di Scienze e Tecnologie Agroambientali (Di.S.T.A.), Patologia Vegetale, Alma Mater Studiorum, Università di Bologna, Italy;(3) Department of Plant Pathology – Plant Molecular Biology/Biotechnology Program, Ohio State University, Columbus, OH, USA;(4) Present address: Department of Molecular Virology, Immunology and MedicanlGenetics/Center for Microbial Interface Biology, Ohio State University, Columbus, OH, USA |
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| Abstract: | A rapid detection method based on PCR amplification of Pseudomonas syringae pv. tomato chromosomal sequences was developed. Primer design was based on the P. syringae DC3000 hrpZPst gene, which maps on a pathogenicity-associated operon of the hrp/hrc pathogenicity island.A 532 bp product corresponding to an internal fragment of hrpZPst was amplified from 50 isolates of P. syringae pv. tomato belonging to a geographically representative collection. The amplification product was also obtained from three coronatine-deficient strains of P. syringae pv. tomato.On the other hand, PCR did not produce any such products from 100 pathogenic and symbiotic bacterial strains of the genera Pseudomonas, Xanthomonas, Erwinia, and Rhizobium and 75 unidentified bacterial saprophytes isolated from tomato plants. The method was tested using leaf and fruit spots from naturally-infected tomato plants and asymptomatic nursery plants and artificially contaminated tomato seeds. The results confirmed the high specificity observed using pure cultures. |
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| Keywords: | diagnostics harpin Lycopersicon esculentum polymerase chain reaction |
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