[Development of Ralstonia solanacearum PCR diagnostics method]
2006
Nikolajchik, E.A. | Potapovich, M.I. | Ignatenko, E.I. | Evtushenkov, A.N., Belarus State Univ., Minsk (Belarus)
Research results of the development of safe and quick method of molecular diagnostics of Ralstonia solanacearum of potato (Solanum tuberosum) based on the polymerase chain reaction (PCR) with the further analysis of restriction fragment length polymorphism (RFLP) were presented. Strains 316 and 317 of Ralstonia solanacearum were used. Four new developed primers (RS11, RS12, RS21 and RS22) corresponded to 16S ribosomal RNA gene sequence and had differences from the sequences of closely related species to forbidden the amplification. Sequence of oligo-nucleotides of RS11, RS12, RS21 and RS22 primers was presented. Principle primers design for the amplification of 16S ribosomal RNA gene of Ralstonia solanacearum was analyzed. Prospective sizes of the fragments in the conditions of restriction by various endonucleases of 16S ribosomal RNA gene of Ralstonia solanacearum strain GMI1000 amplificated with the help of RS11 and RS12 primers were presented. Realized investigation made it possible to prove that PCR-RFLP analysis of the developed RS11, RS12, RS21 and RS22 primers could clearly identificate the collective strains of Ralstonia solanacearum
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