Exopolysaccharide (EPS) synthesis in
Bradyrhizobium japonicum : sequence, operon structure and mutational analysis of an
exo gene cluster
B. U. Becker1, K. Kosch1, M. Parniske1 and P. Müller1
Abstract The nucleotide sequence of a 8330-bp DNA fragment from
Bradyrhizobium japonicum 110
spc4 was determined. Sequence analysis revealed that six ORFs were present and the deduced amino acid sequences were homologous
to enzymes involved in exopolysaccharide (EPS) biosynthesis. The genes appear to be organized into at least four different
operons. One gene was found to be homologous to
exoB, which encodes a UDP-galactose 4′-epimerase. Other ORFs were homologous to UDP-hexose transferases and one ORF showed similarity
to
Sinorhizobium (Rhizobium) meliloti ExoP, which has been suggested to be involved in EPS chain-length determination. A set of deletion and insertion mutants was
constructed and the resulting
B. japonicum strains were tested for their symbiotic traits. Deletion mutant ΔP22, which lacks the C-terminal part of ExoP, the UDP-hexose
transferase ExoT and the N-terminal part of ExoB, shows a delayed nodulation phenotype and induces symptoms of plant defense
reactions; its EPS does not contain galactose and no high molecular weight fraction is synthesized. In contrast, insertion
mutant EH3, which expresses an
exoP gene product that is truncated in its putative periplasmic domain, produced an EPS containing both HMW and LMW fractions.
However, the interaction of EH3 with soybeans was severely perturbed. As a rule, only the initial steps of nodule formation
were observed.
Key words Exopolysaccharide - Symbiosis - Nodule development - Transcriptional reporter - Gene fusion