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Characterization of the Xanthomonas campestris pv. campestris Lipopolysaccharide Substructures Essential for Elicitation of an Oxidative Burst in Tobacco Cells

July 2005 , Volume 18 , Number  7
Pages  674 - 681

Sebastian G. Braun , 1 Andreas Meyer , 1 Otto Holst , 2 Alfred Pühler , 1 and Karsten Niehaus 1

1Fakultät für Biologie, Lehrstuhl für Genetik, Universität Bielefeld, Postfach 100131, D-33501 Bielefeld, Germany; 2Research Center Borstel, Analytical Biochemistry, D-23845 Borstel, Germany

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Accepted 4 March 2005.

The lipopolysaccharides (LPS) of gram-negative bacteria are essential for perception of pathogens by animals and plants. To identify the LPS substructure or substructures recognized by plants, we isolated water-phase (w)LPS from different Xanthomonas campestris pv. campestris mutants and analyzed their sugar content and ability to elicit an oxidative burst in tobacco cell cultures. The different wLPS species are characterized by lacking repetitive subunits of the O-antigen, the complete O-antigen, or even most of the core region. Because loss of lipid A would be lethal to bacteria, pure lipid A was obtained from X. campestris pv. campestris wild-type wLPS by chemical hydrolysis. The elicitation experiments with tobacco cell cultures revealed that LPS detection is dependent on the bioavailability of the amphiphilic wLPS, which can form micelles in an aqueous environment. By adding deoxycholate to prevent micelle formation, all of the tested wLPS species showed elicitation capability, whereas the lipid A alone was not able to trigger an oxidative burst or calcium transients in tobacco cell cultures. These results suggest that the LPS substructure recognized by tobacco cells is localized in the inner core region of the LPS, consisting of glucose, galacturonic acid, and 3-deoxy-d-manno-oct-2-ulosonic acids. Although lipid A alone seems to be insufficient to induce an oxidative burst in tobacco cell cultures, it cannot be ruled out that lipid A or the glucosamine backbone may be important in combination with the inner core structures.

Additional keywords: nonhost resistance , pathogen-associated molecular patterns (PAMPs) .

© 2005 The American Phytopathological Society