DIFFERENTIATION OF PSEUDOMONAS-SOLANACEARUM, PSEUDOMONAS-SYZYGII, PSEUDOMONAS-PICKETTII AND THE BLOOD-DISEASE BACTERIUM BY PARTIAL 16S-RIBOSOMAL-RNA SEQUENCING - CONSTRUCTION OF OLIGONUCLEOTIDE PRIMERS FOR SENSITIVE DETECTION BY POLYMERASE CHAIN-REACTION

The sequence of a 292 bp segment of the DNA encoding 16S rRNA (corresponding to positions 44-337 of the Escherichia coli 16S RRNA sequence) was determined for each of 40 Pseudomonas solanacearum, four banana Blood Disease Bacterium, three P. syzygii and two P. pickettii strains. Phylogenetic relationships derived from comparison of these sequences to each other, and to equivalent 16S rRNA gene sequences from other bacteria present in the EMBL databank, conform well with those obtained previously by DNA-DNA/rRNA hybridization experiments. The 16S rRNA sequence of the Blood Disease Bacterium was identical over the 292 bp to one of the four sequence groups of P. solanacearum, suggesting that these pseudomonads are more closely related to each other than to P. syzygii or P. pickettii. Sequence data comparisons allowed construction of an oligonucleotide specific for P. solanacearum, P. syzygii and the Blood Disease Bacterium. Use of the specific oligonucleotide with a non-specific oligonucleotide in the polymerase chain reaction enabled 1-10 cells of bacteria in this group to be detected after 50 rounds of amplification by visualizing a 287-288 bp product on agarose gels.