P-171 - Detection and comparison of Curtobacterium flaccumfaciens pv. flaccumfaciens (Cff) using seed extract and molecular methods

Bacterial wilt, caused by Curtobacterium flaccumfaciens pv. flaccumfaciens (Cff), has recently been detected in certain regions of Europe in imported seeds from the U.S. The primary hosts of this pathogen are legume species from the Fabaceae family, including common beans and garden beans. To address this issue, the European Union Reference Laboratory for Plant Pathogenic Bacteria (EURL-BAC) has developed a diagnostic procedure for the official testing of common bean (Phaseolus vulgaris) seeds for the presence of Cff, as testing is now mandatory for seed lots before importation. This study aimed to detect Cff in beans seed lots and then perform PCR on suspect colonies to correlate the results. The presence of Cff was verified by plating seed extracts on agar media to isolate potential Cff colonies. Suspect colonies were then subjected to conventional PCR following the method of Tegli et al. (2002) and ISHI-Veg real-time TaqMan PCR, developed by Naktuinbouw, as outlined in the EURL-BAC-2024-CORBFL-TM-01 procedure. Being able to detect Cff is crucial for preventing the spread of the disease. The results were promising, as the PCR findings were successfully correlated.