Sinorhizobium meliloti pSymB carries genes necessary for arabinose transport and catabolism.
Arabinose is a known component of plant cell walls and is found in the rhizosphere. In this work, a previously undeleted region of the megaplasmid pSymB was identified as encoding genes necessary for arabinose catabolism, by Tn5-B20 random mutagenesis and subsequent complementation. Transcription of this region was measured by beta-galactosidase assays of Tn5-B20 fusions, and shown to be strongly inducible by arabinose, and moderately so by galactose and seed exudate. Accumulation of [(3)H]arabinose in mutants and wild-type was measured, and the results suggested that this operon is necessary for arabinose transport. Although catabolite repression of the arabinose genes by succinate or glucose was not detected at the level of transcription, both glucose and galactose were found to inhibit accumulation of arabinose when present in excess. To determine if glucose was also taken up by the arabinose transport proteins, [(14)C]glucose uptake rates were measured in wild-type and arabinose mutant strains. No differences in glucose uptake rates were detected between wild-type and arabinose catabolism mutant strains, indicating that excess glucose did not compete with arabinose for transport by the same system. Arabinose mutants were tested for the ability to form nitrogen-fixing nodules on alfalfa, and to compete with the wild-type for nodule occupancy. Strains unable to utilize arabinose did not display any symbiotic defects, and were not found to be less competitive than wild-type for nodule occupancy in co-inoculation experiments. Moreover, the results suggest that other loci are required for arabinose catabolism, including a gene encoding arabinose dehydrogenase.
Poysti
Nathan J
NJ
Department of Microbiology, University of Manitoba, Winnipeg, MB R3T 2N2, Canada.
Loewen
Erin D M
EDM
Department of Microbiology, University of Manitoba, Winnipeg, MB R3T 2N2, Canada.
Wang
Zexi
Z
Department of Microbiology, University of Manitoba, Winnipeg, MB R3T 2N2, Canada.
Oresnik
Ivan J
IJ
Department of Microbiology, University of Manitoba, Winnipeg, MB R3T 2N2, Canada.
D001089:Arabinose / Q000378:metabolism*; D019677:Artificial Gene Fusion; D001692:Biological Transport; D004251:DNA Transposable Elements; D005690:Galactose / Q000494:pharmacology; D017353:Gene Deletion; D015964:Gene Expression Regulation, Bacterial; D017930:Genes, Reporter; D005816:Genetic Complementation Test; D005947:Glucose / Q000494:pharmacology; D000455:Medicago sativa / Q000382:microbiology; D016254:Mutagenesis, Insertional; D010957:Plasmids / Q000235:genetics*; D053204:Root Nodules, Plant / Q000382:microbiology; D016962:Sinorhizobium meliloti / Q000235:genetics* / Q000378:metabolism; D014158:Transcription, Genetic; D001616:beta-Galactosidase / Q000032:analysis
D004251:DNA Transposable Elements; D001089:Arabinose; D001616:beta-Galactosidase; D005947:Glucose; D005690:Galactose