From 2002 to 2004, tomato (Lycopersicon esculentum) plants with external stem lesions, adventitious roots, and necrotic pith that was hollowed or chambered were received by the Clinical Lab of the Plant Protection Department from eight greenhouses in the Riyadh, Abha, and El-Kharj regions of Saudi Arabia. Bacteria were isolated on nutrient agar or King's medium B (KMB) from the stems of tomato plants cv. Red Gold, the cultivar most commonly grown in greenhouses. Gram-negative, rod-shaped bacteria were consistently isolated from stems with symptoms of pith necrosis. They were identified as Pseudomonas fluorescens (biotype I) and P. corrugata on the basis of morphological, physiological, and biochemical tests (2). Isolates of P. fluorescens isolated from Abha and El-Kharj were fluorescent on KMB, aerobic, and positive for oxidase, arginine dihydrolase, and gelatin liquefaction. Furthermore, all isolates produced levan-type colonies on sucrose nutrient agar and utilized glucose, 2-ketogluconate, sucrose, and sorbitol. They were negative for tobacco hypersensitivity and nitrate reduction. The strains of P. corrugata isolated from Riyadh were nonfluorescent, aerobic, and positive for oxidase, nitrate reductase, arginine dihydrolase, and utilization of malonate, alanine, trehalose, arginine, mannitol, and m-inositol. They were negative for levan, pectinase, tobacco hypersensitivity, and utilization of cellobiose and sorbitol. The identity of bacterial species was confirmed by Biolog analysis (carbon source utilization at 37°C), with a similarity index of 0.75 for P. corrugata and 0.71 for P. fluorescens. Four-week-old tomato plants (cv. Red Gold) were inoculated by injecting 50 μl of a bacterial suspension into the axils of the first true leaves. The bacterial suspension was prepared from 24-h-old cultures with sterile distilled water. Sterile distilled water was used as the negative control. After inoculation, plants were covered with polyethylene bags for 24 h to maintain high humidity at 25°C (1). Necrotic lesions surrounding injection points were observed 14 days after inoculation. At 4 weeks after inoculation, all inoculated plants showed symptoms of necrotic pith similar to those observed on the samples received. Control plants injected with water remained healthy throughout the experiments. Isolates of P. fluorescens (biotype I) and P. corrugata were reisolated from inoculated plants and were identical to the original strains on the basis of Biolog analysis. To our knowledge, this is the first report of tomato pith necrosis in Saudi Arabia. References: (1) G. Demir. J. Turk. Phytopathol. 19:63, 1990. (2) R. A. Lelliott and D. E. Stead. Methods for the Diagnosis of Bacterial Diseases of Plants. Blackwell Scientific Publications. Oxford, UK, 1987.
Pith necrosis associated with Pseudomonas viridiflava in tomato plants in Brazil