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Purification and Visualization of Lipopolysaccharide from Gram-negative Bacteria by Hot Aqueous-phenol Extraction
Published on: May 28, 2012
Differences between lipopolysaccharide compositions of plant pathogenic and saprophytic pseudomonas species
1Department of Biology, Utah State University, Logan, Utah 84322.
Abstract:
Lipopolysaccharides (LPS) were obtained by washing cells of plant pathogenic and saprophytic Pseudomonas species with saline (fraction 1) and then with saline-EDTA (fraction 2). The cells subsequently were extracted with phenol to yield a third aqueous preparation (fraction 3). Each fraction type contained the LPS components, lipid A, heptose, 2-keto-3-deoxy sugar, and neutral and amino sugars. The neutral sugar compositions of fractions 1, 2, and 3, although similar within a species, differed between the Pseudomonas species. The LPS of two pathovars (pv.) of Pseudomonas syringae had glucose and rhamnose as major components: 13 (+/-3)% glucose and 87 (+/-3)% rhamnose for P. syringae pv. pisi and 18 (+/-5)% glucose and 76 (+/-2)% rhamnose for P. syringae pv. syringae. Fucose was present in addition to glucose and rhamnose for P. syringae pv. phaseolicola (68 [+/-8]% rhamnose, 14 [+/-1]% fucose, and 14 [+/-5]% glucose) and P. syringae pv. tabaci (24 [+/-2]% rhamnose, 54 [+/-3]% fucose, and 17 [+/-1]% glucose). The LPS from different races of P. syringae pv. pisi and P. syringae pv. phaseolicola could not be distinguished by neutral sugar composition. Three saprophytic species, P. aeruginosa, P. fluorescens, and P. putida, also produced LPS which had different proportions of rhamnose, fucose, and glucose. The LPS from three isolates of P. putida were distinct in possessing a high proportion of amino sugar and containing glucose as the major neutral sugar component (86 to 100%). The LPS fractions from plant pathogenic and saprophytic Pseudomonas species did not elicit browning or phytoalexin production in treated dark red kidney bean cotyledons or red Mexican bean leaves. Rather, chlorosis of the LPS-treated leaf tissue was observed.
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