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Journal of Bacteriology, October 2000, p. 5317-5324, Vol. 182, No. 19
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Varying the Abundance of O Antigen in Rhizobium etli and Its Effect on Symbiosis with Phaseolus vulgaris

K. Dale Noel,1,* Lennart S. Forsberg,2 and Russell W. Carlson2

Department of Biology, Marquette University, Milwaukee, Wisconsin,1 and Complex Carbohydrate Research Center, University of Georgia, Athens, Georgia2

Received 7 April 2000/Accepted 9 July 2000

Judged by migration of its lipopolysaccharide (LPS) in gel electrophoresis, the O antigen of Rhizobium etli mutant strain CE166 was apparently of normal size. However, its LPS sugar composition and staining of the LPS bands after electrophoresis indicated that the proportion of its LPS molecules that possessed O antigen was only 40% of the wild-type value. Its LPS also differed from the wild type by lacking quinovosamine (2-amino-2,6-dideoxyglucose). Both of these defects were due to a single genetic locus carrying a Tn5 insertion. The deficiency in O-antigen amount, but not the absence of quinovosamine, was suppressed by transferring into this strain recombinant plasmids that shared a 7.8-kb stretch of the R. etli CE3 lps genetic region alpha , even though this suppressing DNA did not carry the genetic region mutated in strain CE166. Strain CE166 gave rise to pseudonodules on legume host Phaseolus vulgaris, whereas the mutant suppressed by DNA from lps region alpha  elicited nitrogen-fixing nodules. However, the nodules in the latter case developed slowly and were widely dispersed. Two other R. etli mutants that had one-half or less of the normal amount of O antigen also gave rise to pseudonodules on P. vulgaris. The latter strains were mutated in lps region alpha  and could be restored to normal LPS content and normal symbiosis by complementation with wild-type DNA from this region. Hence, the symbiotic role of LPS requires near-normal abundance of O antigen and may require a structural feature conferred by quinovosamine.

* Corresponding author. Mailing address: Department of Biology, Marquette University, P.O. Box 1881, Milwaukee, WI 53201-1881. Phone: (414) 288-1475. Fax: (414) 288-7357. E-mail: dale.noel{at}marquette.edu.

Journal of Bacteriology, October 2000, p. 5317-5324, Vol. 182, No. 19
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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