First Report of the Purple Variant of <i>Curtobacterium flaccumfaciens</i> pv. <i>flaccumfaciens</i>, Causal Agent of Bacterial Wilt of Bean, in Canada

Bacterial wilt of bean (Phaseolus vulgaris L.) caused by the yellow and orange variants of Curtobacterium flaccumfaciens pv. Flaccumfaciens (Hedges) Collins & Jones was found in western Canada in 2002 (1). A purple variant was found in a pooled sample of discolored cull seeds of great northern bean (cv. US1140) from a crop grown near Bow Island, Alberta, Canada in 2005. Bacterial colonies isolated from purple seed using modified Burkholder's agar (MBA) (3) were convex, glistening, and smooth edged with blue pigment diffusing into the medium. Three isolates (V154, V155, and V254) were identified with conventional tests (2), carbohydrate oxidation (GP Microplates, Biolog Inc., Hayward, CA), and cellular fatty acids (CFA) (MIDI, Inc., Newark, DE). All were grampositive, motile, aerobic rods with yellow colonies producing extracellular blue pigment on MBA when grown at 20 ± 2°C. Bacterial isolates grew at 27°C but grew weakly at 37°C. They were positive for catalase and hydrolysis of hippurate and indoxyl acetate and negative for urease, gelatin liquification, and oxidase. CFA profiles were approximately 48% 15:0 anteiso, 40% 17:0 anteiso, 7% 16:0 iso, and 3% 15:0 iso; with 17:1 anteiso A variable but <1%. Many carbohydrates were oxidized in the Biolog microplates with little acid production. The results match C. flaccumfaciens (2) and the MIDI and Biolog databases, as well as the purple variant of C. flaccumfaciens found in Nebraska, the only previous report of this variant (4). The pathogenicity of the three isolates was tested. Seeds of great northern (cv. US1140) and navy (cv. Morden003) beans were soaked in a bacterial suspension (1 × 108 CFU/ml) or distilled water (control) for 1 h, planted in Cornell mix in root trainers, incubated at 28/22°C (16-h day/8-h night) in a growth cabinet for 14 days, and examined for seedling wilt. The test had three replicates per treatment and 20 seeds per replicate in a completely randomized design. All three isolates were pathogenic to both bean cultivars. The wilt incidences were 51, 57, and 56% on US1140 and 64, 76, and 69% on Morden003 for isolates V154, V155, and V254, respectively. The purple variant of C. flaccumfaciens was reisolated from hypocotyls of wilted seedlings but not from healthy controls. The experiment was repeated using the reisolated bacteria and the results were similar to the first experiment, thus fulfilling Koch's postulates. To our knowledge, this is the first report of the purple variant of C. flaccumfaciens pv. flaccumfaciens in Canada. References: (1) T. F. Hsieh et al. Plant Dis. 86:1275, 2002. (2) K. Komagata et al. Page 1313 in: Bergey's Manual of Systematic Bacteriology. Vol. 2. Williams and Wilkens, Baltimore, MD, 1986. (3) G. A. Nelson and G. Semeniuk. Phytopathology 54:330, 1964. (4) M. L. Schuster et al. Can. J. Microbiol. 14:423, 1968.

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