Abstract
Summary PCR patterns of five nitrogen-fixing soil bacterial strains isolated from the rhizosphere of wheat in the vicinity of Bayreuth were generated by PCR using random and repetitive (BOX, ERIC and REP) primers. They have been compared to the patterns (obtained with the same primers) of several Rahnella aquatilis strains (including the type strain of this species - R. aquatilis ATCC 33071), the strain R. aquatilis ATCC 33989 (to whom one of our isolates has been formerly assumed to be related), and two clinical isolates of the same species. As outgroup strains Enterobacter agglomerans 19-78, Escherichia coli W2438 and Pantoea agglomerans 5D representing different species of the family Enterobacteriaceae were used. By all primers used it was clearly shown that the natural isolates belong to the species R. aquatilis . Even more, they have been clustered into two groups around the two ATCC strains 33071 and 33989, respectively. By these analyses, it was possible also to show that the clinical R. aquatilis strains form another, third group. In addition, both kinds of PCR fingerprinting (using arbitrary or repetitive primers) generated highly reproducible patterns when parallel reactions with total DNA extracted by different methods from independent liquid cultures of one and the same strain were performed. The patterns obtained by PCR fingerprinting of total DNA or of cells from fresh colonies or liquid cultures added to the PCR mixture did not differ significantly. The RAPD and rep-APD characterization of the strains studied here is in full agreement with their taxonomical analysis performed by other molecular methods such as micro- and macro-RFLP fingerprinting, ribotyping and 16S rDNA sequencing. On the basis of these results we recommend to apply these simple, fast and cheap methods for identification and discrimination of new environmental and clinical isolates of the species R. aquatilis.
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