The development of a specific Real-Time TaqMan for the detection of Clavibacter michiganensis supsp. michiganensis
J. OOSTERHOF (1), S. Berendsen (1)
(1) Rijk Zwaan Breeding BV, DeLier, NETHERLANDS
Phytopathology 101:S133

The detection of Clavibacter michiganensis supsp. michiganensis is of great economical importance. This seed born pathogen, with quarantine status, is the causative agent of bacterial cancer in tomato and causes economic losses in commercial tomato and seed production. The currently used detection methods all have drawbacks. The development of a new detection method was therefore still needed. ISHI-NL used AFLP^® to find specific fragments present in all selected Clavibacter michiganensis supsp. michiganensis isolates. Only one single fragment of interest was found. The fragment was partial coding for a gene producing a protein two-component system sensor kinase. This gene was used to develop the specific RZ_Ptssk MGB based TaqMan^®. The developed RZ_Ptssk TaqMan^® was tested and compared with existing Real-Time PCR assay’s on a collection of 67 isolates. The RZ_Ptssk TaqMan was the only assay correlating 100% with a pathogenicity test on tomato plants. Several ISHI_NL and international labs are using the RZ_Ptssk TaqMan^® for their own specific purpose. Until today no false negative or false positive isolates have been reported.