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Journal of Bacteriology, October 2001, p. 6054-6064, Vol. 183, No. 20
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.20.6054-6064.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Genetic Locus Required for Antigenic Maturation of Rhizobium etli CE3 Lipopolysaccharide

Dominik M. Duelli,1,dagger Andrea Tobin,2 Jodie M. Box,1 V. S. Kumar Kolli,2 Russell W. Carlson,2 and K. Dale Noel1,*

Department of Biology, Marquette University, Milwaukee, Wisconsin 53233,1 and Complex Carbohydrate Research Center, The University of Georgia, Athens, Georgia 306022

Received 16 January 2001/Accepted 6 August 2001

Rhizobium etli modifies lipopolysaccharide (LPS) structure in response to environmental signals, such as low pH and anthocyanins. These LPS modifications result in the loss of reactivity with certain monoclonal antibodies. The same antibodies fail to recognize previously isolated R. etli mutant strain CE367, even in the absence of such environmental cues. Chemical analysis of the LPS in strain CE367 demonstrated that it lacked the terminal sugar of the wild-type O antigen, 2,3,4-tri-O-methylfucose. A 3-kb stretch of DNA, designated as lpe3, restored wild-type antigenicity when transferred into CE367. From the sequence of this DNA, five open reading frames were postulated. Site-directed mutagenesis and complementation analysis suggested that the genes were organized in at least two transcriptional units, both of which were required for the production of LPS reactive with the diagnostic antibodies. Growth in anthocyanins or at low pH did not alter the specific expression of gusA from the transposon insertion of mutant CE367, nor did the presence of multiple copies of lpe3 situated behind a strong, constitutive promoter prevent epitope changes induced by these environmental cues. Mutations of the lpe genes did not prevent normal nodule development on Phaseolus vulgaris and had very little effect on the occupation of nodules in competition with the wild-type strain.

* Corresponding author. Mailing address: Department of Biology, Marquette University, Milwaukee, WI 53233. Phone: (414) 288-1475. Fax: (414) 288-7357. E-mail: Dale.Noel{at}marquette.edu.

dagger Present address: Cold Spring Harbor Laboratory, Cold Spring Harbor, NY 11724.

Journal of Bacteriology, October 2001, p. 6054-6064, Vol. 183, No. 20
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.20.6054-6064.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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