Genetic relationships among strains of Xanthomonas fragariae based on random amplified polymorphic DNA PCR, repetitive extragenic palindromic PCR, and enterobacterial repetitive intergenic consensus PCR data and generation of multiplexed PCR primers useful for the identification of this phytopathogen

• Published in: Applied and Environmental Microbiology Vol. 62; no. 9; pp. 3121 - 3127
• Main Authors: Pooler, M.R. (U.S. National Arboretum, Washington, DC.), Ritchie, D.F, Hartung, J.S
• Format: Journal Article
• Language: English
• Published: Washington, DC American Society for Microbiology 01.09.1996
• Subjects: ADN; Bacteria; Bacterial plant pathogens; Base Sequence; Biological and medical sciences; CHIMIOTAXONOMIE; DIFERENCIAS BIOLOGICAS; DIFFERENCE BIOLOGIQUE; DNA Primers; DNA, Bacterial - analysis; Fundamental and applied biological sciences. Psychology; Genetics; GENOTIPOS; GENOTYPE; IDENTIFICACION; IDENTIFICATION; Microbiology; Molecular Sequence Data; PCR; Phytopathology. Animal pests. Plant and forest protection; POLIMORFISMO GENETICO; Polymerase Chain Reaction; POLYMORPHISME GENETIQUE; QUIMIOTAXONOMIA; Systematics. Structure, properties and multiplication. Genetics; XANTHOMONAS; Xanthomonas - genetics; Xanthomonas - pathogenicity; Xanthomonas - physiology; Xanthomonas fragariae; Pseudomonadales; Genetic variability; Genetic marker; Plant pathogen; Primer; Genetic distance; Identification; Method; Polymerase chain reaction; Random amplified polymorphic DNA; Bacteria; Pseudomonadaceae; Xanthomonas fragariae; Polymorphism
• ISSN: 0099-2240; 1098-5336
• DOI: 10.1128/aem.62.9.3121-3127.1996

Genetic relationships among 25 isolates of Xanthomonas fragariae from diverse geographic regions were determined by three PCR methods that rely on different amplification priming strategies: random amplified polymorphic DNA (RAPD) PCR, repetitive extragenic palindromic (REP) PCR, and enterobacterial repetitive intergenic consensus (ERIC) PCR. The results of these assays are mutually consistent and indicate that pathogenic strains are very closely related to each other. RAPD, ERIC, and REP PCR assays identified nine, four, and two genotypes, respectively, within X. fragariae isolates. A single nonpathogenic isolate of X. fragariae was not distinguishable by these methods. The results of the PCR assays were also fully confirmed by physiological tests. There was no correlation between DNA amplification product patterns and geographic sites of isolation, suggesting that this bacterium has spread largely through exchange of infected plant germ plasm. Sequences identified through the RAPD assays were used to develop three primer pairs for standard PCR assays to identify X. fragariae. In addition, we developed a stringent multiplexed PCR assay to identify X. fragariae by simultaneously using the three independently derived sets of primers specific for pathogenic strains of the bacteria