Isolation and characterization of lipopolysaccharides from different rhizobial isolates

Formation of nodules on roots or in stems (in some cases) of leguminous plants is the unique ability of gram-negative bacteria, Rhizobia, which converts atmospheric nitrogen into usable forms by the host plant. Lipopolysaccharide (LPS) is the outer membrane component of the gram-negative bacteria, known to be an essential factor in host recognition, specificity, and initial infection processes. In the present study, we extracted lipopolysaccharides from different rhizobial isolates by a modified phenol-water method and partially characterized by polyacrylamide gel electrophoresis with silver staining. The results showed two separate banding regions, LPS-I and LPS-II. The high molecular weight and electrophoretic mobility of LPS-I region resembles that of lysozyme, used as a standard marker. The LPS-II region has a low molecular weight and electrophoretic mobility greater than that of lysozyme. The LPS-II region was due to incomplete LPS, which either lacks the entire O-antigen repeating unit or contains only one or two repeating units. The banding patterns of LPS vary among the different rhizobial isolates. Results revealed that the type of LPS structure and banding regions greatly facilitate the further characterization of the LPS modifications required for symbiosis.