Detection: Ralstonia solanacearum on potato.
Matrix: Bacterial colonies isolated from potato tuber tissue or directly in potato tuber tissue. Sampling and sample preparation are described in the PM7/21 (2) from EPPO.
Other information: set developed and optimized by Qualiplante according to Weller et al., 2000. A verification was performed by Qualiplante (data not published) and the performance characteristics of the set are the same as the original publication.
The broad-host-range R. solanacearum probe (RS-P) is partially homologous to 16S rRNA gene primer OLI1 (24), with primers (RS-I and RS-II) flanking this region. This set permits the detection of all strains of R. solanacearum and R. pseudosolanacearum and the specific detection and identification of R. solanacearum Phylotype II sequevar 1 (race 3) only. The broad-range probe (RS-P) detected all biovars of R. Solanacearum.
In pure culture, detection of Ralstonia solanacearum to ≥ 10-2 cells ml-1 was achieved; sensitivity decreased when the assay was performed with inoculated potato tissue extracts, prepared by currently recommended extraction procedures.
No cross reactions were observed with Ralstonia pickettii, Ralstonia sp., Burkholderia andropogonis, Burkholderia caryophylli, Burkholderia cepacia, Burkholderia glumae, Burkholderia plantarii, Bacilus polymyxa, Pseudomonas marginalis subsp. marginalis, Pseudomonas chlorophis, Enterobacteriaceae, Ratinella aquatillis, Ochrobactrum anthropic.
Cross reactions were observed with Banana blood disease bacterium, Ralstonia syzygii strains, Pseudomonas sp. (Taxom B).
This method is referred in the Appendix 5 of the PM 7/21 (2) Ralstonia solanacearum, R. pseudosolanacearum and R. syzygii (Ralstonia solanacearum species complex), European and Mediterranean Plant Protection Organization Bulletin (2018) 48 (1), 32-63.
Taq-Man® technology, FAM fluorophore.
Included reagents: Direct Master Mix (DM), Negative Control (NC), Positive Control (PC).