Characterisation of Rhizobium isolates by amplification of DNA polymorphisms using random primers

Abstract

The use of single random primers, selected in the absence of target sequence information, has been shown to be effective in producing DNA amplifications that provide fingerprints which are unique to individual organisms. DNA amplification by random priming was applied to the DNA from isolates of Rhizobium leguminosarum biovar trifolii. Amplification products were produced using a number of primers, and the resulting fingerprints allowed strain differentiation. However, the effectiveness of primers was dependent upon length and GC content. It was also possible to amplify DNA directly from cells in culture and in nodule tissue. Lysis of these cells was achieved simply through heat applied in the initial DNA denaturation stage of the thermal reaction. The ability to produce varied amplification patterns from different Rhizobium isolates, especially directly from nodules, gives this method potential for use in examining genetic structures and relationships in Rhizobium populations. Key words: Rhizobium, DNA amplification, random primers.