Degree Name
Master of Science (MS)
Semester of Degree Completion
1990
Thesis Director
Kandy D. Baumgardner
Abstract
The lipopolysaccharides (LPSs) from Rhizobium leguminosarum biovar trifolii TB4 and two transposon (Tn5) symbiotic mutants, R. leguminosarum bv. trifolii TB104 and TB112, were isolated and partially characterized. Bv. trifolii TB4 is a derivative of R. leguminosarum bv. trifolii ANU843. The two mutant strains elicited incompletely developed clover nodules that exhibited low bacterial populations and very low nitrogenase activity. The LPSs were extracted using the hot phenol-water method and purified with gel filtration column chromatography using Sepharose 4B with an elution buffer consisting of EDTA and triethylamine. The hexose compositions were determined by gas chromatography of the alditol acetate derivatives. LPS components were quantitated by colorimetric assays. The LPSs of both mutants lacked O-antigen sugars and some sugar residues of the LPS core oligosaccharides. Therefore, R. leguminosarum bv. trifolii TB104 and TB112 are missing LPS I, the complete form of LPS containing the O-antigen found in the parent strain TB4. Polyacrylamide gel electrophoresis analysis verifies that the mutants are missing the O-antigen through the lack of bands in the LPS I higher molecular weight region. An immunoblot procedure using bv. trifolii ANU843 antisera confirmed these results. Thus, while the parent strain contains both LPS I (O-antigen complete) and LPS II (O-antigen incomplete), the mutants contain only LPS II.
Recommended Citation
Miller, Jill K., "The Isolation and Partial Characterization of the Lipopolysaccharides from Rhizobium leguminosarum biovar trifolii strains TB4, TB104, and TB112" (1990). Masters Theses. 2302.
https://thekeep.eiu.edu/theses/2302
