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A diagnostic medium for the semi-selective isolation and enumeration of <Emphasis Type="Italic">Xanthomonas axonopodis</Emphasis>pv. <Emphasis Type="Italic">vignicola</Emphasis>
Authors:Kerstin Wydra  Gopal Khatri-chhetri  Athanassios Mavridis  Rachidatou Sikirou  Klaus Rudolph
Institution:(1) Institute of Plant Pathology and Plant Protection, University of Göttingen, Grisebachstr. 6, Göttingen, 37077, Germany;(2) International Institute of Tropical Agriculture, B.P. 08 Tri Postal, 0932 Cotonou, Benin;(3) Institute of Plant Diseases and Plant Protection, University of Hannover, Herrenhäuser Str. 2, Hannover, 30419, Germany
Abstract:A semi-selective medium for isolation of Xanthomonas axonopodis pv. vignicola from cowpea (Vigna unguiculata) plant and soil samples was developed. Twelve carbon and five nitrogen sources were tested with four strains of X. axonopodispv.vignicola, and 25 antibiotics were screened against saprophytes. -cellobiose (10g) was selected as the optimal carbon source. Among the antibiotics, cefazoline inhibited growth of most of the saprophytes with little effect on strains of the pathogen. ,-methionine enhanced growth of X. axonopodispv.vignicola. Boric acid along with ammonium chloride suppressed growth of Pseudomonas fluorescens. The semi-selective medium designated as cefazoline-cellobiose-methionine (CCM) medium contained K2HPO4 1.34g, KH2PO4 0.4g, MgSO4 0.3g, H3BO3 0.2g, NH4Cl 1.0g, -cellobiose 10g, cycloheximide 0.2g, ,-methionine 1.0g, cefazoline 10mg and agar 14g per l of water (pH 7.2). Colonies of X. axonopodispv.vignicola on CCM medium were whitish, round, raised and 0.2–1.8mm in diameter 96h after incubation. CCM medium generally inhibited growth of Pantoea agglomerans, Bacillus subtilis and saprophytes isolated from cowpea leaves. Colonies of Pseudomonas fluorescens and a saprophytic bacterium, which were not completely suppressed by CCM, could be differentiated from X. axonopodispv.vignicola by their smaller size and different color. The CCM medium proved useful for isolation of X. axonopodispv.vignicola from cowpea plant and soil samples. This is the first report of a semi-selective medium developed for detection of X. axonopodispv.vignicola.
Keywords:cefazoline cellobiose methionine medium  Vigna unguiculata  Xanthomonas campestrispv  vignicola
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