Detection and identification of phytopathogenic Xanthomonas strains by amplification of DNA sequences related to the hrp genes of Xanthomonas campestris pv. vesicatoria

• Published in: Applied and Environmental Microbiology Vol. 60; no. 4; pp. 1068 - 1077
• Main Authors: Leite, R.P. Jr, Minsavage, G.V, Bonas, U, Stall, R.E
• Format: Journal Article
• Language: English
• Published: Washington, DC American Society for Microbiology 01.04.1994
• Subjects: ADN; AGENT PATHOGENE; AMPLIFICATION CHAINE POLYMERASE; BACTERIA; Bacteriology; Base Sequence; Biological and medical sciences; Biology; Deoxyribonucleic acid; DNA; DNA, Bacterial - genetics; Fundamental and applied biological sciences. Psychology; GENE; GENES; Genes, Bacterial; Genetics; IDENTIFICACION; IDENTIFICATION; Microbiology; Molecular Sequence Data; Nucleic Acid Hybridization; Oligonucleotide Probes; ORGANISMOS PATOGENOS; PATHOTYPE; PATOTIPOS; Plant Diseases - microbiology; PODER PATOGENO; Polymerase Chain Reaction; POUVOIR PATHOGENE; REACCION DE CADENAS DE POLIMERASA; Sequence Homology, Nucleic Acid; Species Specificity; Virulence - genetics; XANTHOMONAS; Xanthomonas - classification; Xanthomonas - genetics; Xanthomonas - isolation & purification; Xanthomonas - pathogenicity; XANTHOMONAS CAMPESTRIS; Xanthomonas campestris - genetics; Xanthomonas campestris - isolation & purification; Xanthomonas campestris - pathogenicity; Pseudomonadales; Amplification; Molecular hybridization; Xanthomonas campestris; Gene cluster; Genome organization; Plant pathogen; Nucleotide sequence; Bacteria; Pseudomonadaceae; Molecular probe; Detection
• ISSN: 0099-2240; 1098-5336
• DOI: 10.1128/aem.60.4.1068-1077.1994

Three pairs of oligonucleotide primers specific for different regions of the hrp gene (hypersensitive reaction and pathogenicity) cluster of Xanthomonas campestris pv. vesicatoria were designed and tested for amplification of DNA isolated from a large number of different bacteria. DNA sequences related to the hrp genes were successfully amplified from X. fragariae and from 28 pathovars of X. campestris. No DNA amplification occurred with genomic DNA from phytopathogenic strains of X. campestris pv. secalis, X. campestris pv. translucens, and X. albilineans or from nonpathogenic opportunistic xanthomonads and phytopathogenic strains of the genera Acidovorax, Agrobacterium, Clavibacter, Erwinia, Pseudomonas, and Xylella. The DNA from those bacteria also failed to hybridize to hrp-specific fragments in Southern blot analysis. DNA fragments amplified with a particular primer pair were of identical size from each of the different phytopathogenic xanthomonads. However, restriction analysis of these fragments by using frequently cutting endonucleases revealed variation in the pattern for these hrp-related fragments amplified from the different Xanthomonas strains. The restriction patterns generated for the different fragments allowed distinction of the strains representing a pathovar or species of phytopathogenic xanthomonads. We believe that DNA amplification with hrp-specific oligonucleotide primers is a highly sensitive and specific method that can be applied for detection and identification of phytopathogenic xanthomonads