Clavibacter michiganensis subsp. michiganensis, Bacterial Canker of Tomato: 1. Conventional and Molecular Identification

In May 2006, tomato leaves with watersoaked areas between leaf veins were detected in glasshouses in Padinska Skela. The early-stage symptoms were followed by rapid desiccation to white and pale brown necrosis and subsequently by wilting of entire tomato plants. In June 2006, symptoms of bacterial canker and wilt were also recorded in manygreenhouses in the Jablanica County in southern Serbia.Samples of diseased tomato plants were collected from several tomato-growing regions in Serbia in order to identify the causal agent using conventional identification methods (isolation, pathogenicity and bacteriological determinative tests). Another objectiveof this study was to confirm the identity of isolated bacterial strains by polymerase chain reaction (PCR method).According to the morphological characteristics observed on NA and NBY media, biochemical characteristics, hypersensitive response in four-o’clock plant leaves and pathogenicity test on tomato seedlings, the investigated strains were identified as C. michiganensis subsp. michiganensis. To confirm the identity of isolated strains two PCR protocols were used. Amplification of expected length DNA fragments, 614 bp and 270 bp, respectively, confirmed that the investigated strains belonged to C. michiganensis subsp. michiganensis