Lipid A and O‐chain modifications cause Rhizobium lipopolysaccharides to become hydrophobic during bacteroid development

• Published in: Molecular microbiology Vol. 39; no. 2; pp. 379 - 392
• Main Authors: Kannenberg, Elmar L., Carlson, Russell W.
• Format: Journal Article
• Language: English
• Published: Oxford, UK Blackwell Science Ltd 01-01-2001; Blackwell Publishing Ltd
• Subjects: Bacterial Adhesion; Chromatography - methods; Culture Media; Electrophoresis, Polyacrylamide Gel; Hydrocarbons; lipid A; Lipid A - chemistry; Lipid A - metabolism; Lipopolysaccharides - chemistry; Lipopolysaccharides - isolation & purification; Lipopolysaccharides - metabolism; O Antigens - chemistry; O Antigens - metabolism; Oxygen - pharmacology; Pisum sativum - microbiology; Pisum sativum - physiology; Rhizobium leguminosarum; Rhizobium leguminosarum - chemistry; Rhizobium leguminosarum - genetics; Rhizobium leguminosarum - growth & development; Rhizobium leguminosarum - metabolism; Surface Properties
• ISSN: 0950-382X; 1365-2958
• DOI: 10.1046/j.1365-2958.2001.02225.x

Modifications to the lipopolysaccharide (LPS) structure caused by three different growth conditions were investigated in the pea‐nodulating strain Rhizobium leguminosarum 3841. The LPSs extracted by hot phenol–water from cultured cells fractionated into hydrophilic water and/or hydrophobic phenol phases. Most of the LPSs from cells grown under standard conditions extracted into the water phase, but a greater proportion of LPSs were extracted into the phenol phase from cells grown under acidic or reduced‐oxygen conditions, or when isolated from root nodules as bacteroids. Compared with the water‐extracted LPSs, the phenol‐extracted LPSs contained greater degrees of glycosyl methylation and O‐acetylation, increased levels of xylose, glucose and mannose and increased amounts of long‐chain fatty acids attached to the lipid A moiety. The water‐ and phenol‐phase LPSs also differed in their reactivity with monoclonal antibodies and in their polyacrylamide gel electrophoretic banding patterns. Phenol‐extracted LPSs from rhizobia grown under reduced‐oxygen conditions closely resembled the bulk of LPSs isolated from pea nodule bacteria (i.e. mainly bacteroids) in their chemical properties, reactivities with monoclonal antibodies and extraction behaviour. This finding suggests that, during symbiotic bacteroid development, reduced oxygen tension induces structural modifications in LPSs that cause a switch from predominantly hydrophilic to predominantly hydrophobic molecular forms. Increased hydrophobicity of LPSs was also positively correlated with an increase in the surface hydrophobicity of whole cells, as shown by the high degree of adhesion to hydrocarbons of bacterial cells isolated from nodules or from cultures grown under low‐oxygen conditions. The implications of these LPS modifications are discussed for rhizobial survival and function in different soil and in planta habitats.