Detection of Ralstonia solanacearum in potato tubers by polymerase chain reaction

A new polymerase chain reaction (PCR) assay was developed for the detection of Ralstonia solanacearum in potato tubers. The designed primers PS-1/PS-2 based on the sequence data of the 16S rRNA gene. Using the optimized PCR protocol, it was possible to detect R. solanacearum cells artificially added to concentrated potato extracts in the range of 1-10 colony-forming units (CFU) per PCR reaction mixture (10-100 CFU/ml potato homogenate). No amplification products were obtained, when bacteria belonging to other species or genera were submitted to PCR under the same conditions. A total of 10 different DNA extraction methods were adapted for the isolation of R. solanacearum DNA from potato homogenates and were compared for their suitability as pre-PCR procedures.