First Report of Bacterial Canker of Pepper in Ohio.

Light brown, raised lesions were observed on the leaves of bell pepper (Capsicum annuum L.) plants throughout a commercial field in northwest Ohio in 1999. Bacterial streaming from the lesions was observed microscopically. Five representative, pale yellow colonies isolated on yeast dextrose carbonate medium were selected and purified. All isolates induced a hypersensitive response in Mirabilis jalapa L. plants 24 h after inoculation with a 1 × 108 CFU/ml bacterial suspension. All five were identified as Clavibacter michiganensis subsp. michiganensis by fatty acid methyl ester analysis (mean similarity index [S.I.] = 0.76; MIDI, Newark, DE). Identity was confirmed at the University of Hawaii (W. Kaneshiro and A. Alvarez) by carbon substrate utilization pattern (mean S.I. = 0.75; Biolog, Hayward, CA) and positive reactions with C. michiganensis subsp. michiganensis-specific monoclonal antibodies (clones 103-142-1-1 and 103-148-2-1) in ELISA. DNA extracted from lesions and pure cultures was used as template in a polymerase chain reaction (PCR) assay with primers specific for C. michiganensis subsp. michiganensis (CMM-5 and CMM-6) (1). DNA from a known strain of C. michiganensis subsp. michiganensis served as a positive control, while water and DNA from healthy tomato plants were used as negative controls. A 0.6-kb PCR product was amplified from lesions, pure cultures of all five strains, and positive control DNA, but not from the negative controls. Pathogenicity tests were performed twice on 5- to 6-week-old bell pepper (cvs. Collossal, Lafayette, King Arthur, Brigadier, and Commandant) and tomato (cv. Peto 696) plants. Pepper plants were inoculated with each strain by clipping the lowest petioles with scissors that had been dipped into a bacterial suspension (1 × 108 CFU/ml) or by spray inoculation (approximately 5 ml/plant). Tomato plants were inoculated by clipping. Both inoculation methods included a water control. All five strains caused water-soaked lesions on leaves of all pepper varieties within 7 days after spray inoculation. Pepper plants inoculated by clipping did not develop symptoms. DNA extracts from lesions of challenged pepper plants were positive in PCR. All inoculated tomato seedlings exhibited wilting, streaks, and cankers in the stems and necrosis of leaf margins within 15 days after inoculation. None of the control plants developed symptoms. All five strains were re-isolated from inoculated tomato and pepper plants. The original pepper strains and the strains re-isolated from tomatoes were compared using rep-PCR with ERIC primers (4). DNA fingerprints of the re-isolated strains were identical to those of the original strains and were characteristic of C. michiganensis subsp. michiganensis type C. Bacterial canker is a common disease of tomatoes worldwide and has occurred in Ohio for at least 70 years. However, this is the first report of C. michiganensis subsp. michiganensis infecting peppers in Ohio. While the pathogen does not appear to cause systemic disease in peppers, it may serve as a source of inoculum for tomatoes, which are highly susceptible to the disease and often produced in the same greenhouse as peppers or planted in adjacent fields. Bacterial canker has been reported previously from commercial pepper fields in California (2) and Indiana (3). References: (1) J. Dreier et al. Phytopathology 85:462, 1995. (2) M. Lai. Plant Dis. Rep. 60:339, 1976. (3) R. Latin et al. Plant Dis. 79:860, 1995. (4) F. J. Louws et al. Appl. Environ. Microbiol. 60:2286, 1994.