rep-PCR-mediated genomic fingerprinting: a rapid and effective method to identify Clavibacter michiganensis

• Published in: Phytopathology Vol. 88; no. 8; pp. 862 - 868
• Main Authors: Louws, F.J. (MSU-DOE Plant Research Laboratory, East Lansing.), Bell, J, Medina-Mora, C.M, Smart, C.D, Opgenorth, D, Ishimaru, C.A, Hausbeck, M.K, Bruijn, F.J. de, Fulbright, D.W
• Format: Journal Article
• Language: English
• Published: St. Paul, MN American Phytopathological Society 01.08.1998
• Subjects: Biological and medical sciences; BIOLOGICAL DIFFERENCES; CLAVIBACTER MICHIGANENSIS; CLAVIBACTER MICHIGANENSIS SUBSP. INSIDIOSUM; CLAVIBACTER MICHIGANENSIS SUBSP. MICHIGA; CLAVIBACTER MICHIGANENSIS SUBSP. NEBRASK; CLAVIBACTER MICHIGANENSIS SUBSP. TESSELARIUS; DIFERENCIAS BIOLOGICAS; DIFFERENCE BIOLOGIQUE; DNA FINGERPRINTING; EMPREINTE ADN; EPIDEMIOLOGIA; EPIDEMIOLOGIE; EPIDEMIOLOGY; Fundamental and applied biological sciences. Psychology; Fungal plant pathogens; Generalities. Techniques; GENETIC POLYMORPHISM; GENOMAS; GENOME; GENOMES; HUELLAS GENETICAS ADN; HYPOVIRULENCE; IDENTIFICACION; IDENTIFICATION; Microbiology; Mycological methods and techniques used in mycology; Mycology; NUCLEOTIDE SEQUENCE; PATHOGENICITY; PATOGENICIDAD; PCR; Phytopathology. Animal pests. Plant and forest protection; POLIMORFISMO GENETICO; POLYMERASE CHAIN REACTION; POLYMORPHISME GENETIQUE; POUVOIR PATHOGENE; SECUENCIA NUCLEOTIDICA; SEQUENCE NUCLEOTIDIQUE; STRAIN DIFFERENCES; VIRULENCE; Plant pathogen; Taxonomy; Repeated sequence; Identification; Method; Differential diagnostic; Polymerase chain reaction; Specificity; Clavibacter michiganensis; Fingerprint method; Bacteria; Actinomycetes; Detection
• ISSN: 0031-949X; 1943-7684
• DOI: 10.1094/PHYTO.1998.88.8.862

The genomic DNA fingerprinting technique known as repetitive-sequence-based polymerase chain reaction (rep-PCR) was evaluated as a tool to differentiate subspecies of Clavibacter michiganensis, with special emphasis on C. michiganensis subsp. michiganensis, the pathogen responsible for bacterial canker of tomato. DNA primers (REP, ERIC, and BOX), corresponding to conserved repetitive element motifs in the genomes of diverse bacterial species, were used to generate genomic fingerprints of C. michiganensis subsp. michiganensis, C. michiganensis subsp. sepedonicus, C. michiganensis subsp. nebraskensis, C. michiganensis subsp. tessellarius, and C. michiganensis subsp. insidiosum. The rep-PCR-generated patterns of DNA fragments observed after agarose gel electrophoresis support the current division of C. michiganensis into five subspecies. In addition, the rep-PCR fingerprints identified at least four types (A, B, C, and D) within C. michiganensis subsp. michiganensis based on limited DNA polymorphisms; the ability to differentiate individual strains may be of potential use in studies on the epidemiology and host-pathogen interactions of this organism. In addition, we have recovered from diseased tomato plants a relatively large number of naturally occurring avirulent C. michiganensis subsp. michiganensis strains with rep-PCR fingerprints identical to those of virulent C. michiganensis subsp. michiganensis strains