The use of subtractive hybridization to obtain a DNA probe specific for Pseudomonas solanacearum race 3.
Pseudomonas solanacearum, the causal agent of bacterial wilt, has been classified into three races based on host range and into five biovars based on physiological properties. Strains of race 3 belong exclusively to biovar 2 and primarily affect potatoes. Although this race is thought to have originated in the Andean highlands, it has unusual physiological properties that make it a potential threat to potatoes grown at the cooler latitudes worldwide. Consequently, there is need for a rapid and sensitive method for detection of race 3 strains. We have used subtractive hybridization to enrich for race 3-specific DNA sequences in total race 3 genomic DNA, and thereby obtained a 2 kb clone homologous to DNA from all 28 race 3 strains tested, but with only five of 90 non-race 3 strains. In addition, two larger regions of the genome, containing a minimum of 23 kb of DNA, are also specific for race 3. Deletion of this DNA did not affect virulence. This race 3-specific DNA is a potentially useful diagnostic tool for the detection of race 3 strains.
Cook
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Department of Plant Pathology, University of Wisconsin-Madison 53706.
Sequeira
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D015139:Blotting, Southern; D015342:DNA Probes; D004269:DNA, Bacterial / Q000235:genetics; D005798:Genes, Bacterial; D007612:Kanamycin / Q000494:pharmacology; D009693:Nucleic Acid Hybridization; D010957:Plasmids; D011549:Pseudomonas / Q000235:genetics* / Q000472:pathogenicity; D015183:Restriction Mapping; D013045:Species Specificity; D014170:Transformation, Genetic; D014774:Virulence
D015342:DNA Probes; D004269:DNA, Bacterial; D007612:Kanamycin