A new genetic locus in Sinorhizobium meliloti is involved in stachydrine utilization

• Published in: Applied and Environmental Microbiology Vol. 64; no. 10; pp. 3954 - 3960
• Main Authors: Phillips, D.A. (University of California, Davis, CA.), Sande, E.S, Vriezen, J.A.C, De Bruijn, F.J, Le Rudulier, D, Joseph, C.M
• Format: Journal Article
• Language: English
• Published: Washington, DC American Society for Microbiology 01.10.1998
• Subjects: Agronomy. Soil science and plant productions; Bacteria; Bacteriology; Biochemistry and biology; Biological and medical sciences; Chemical, physicochemical, biochemical and biological properties; Fundamental and applied biological sciences. Psychology; GENBANK/AF016307; Genes; Genetics; LOCI; LOCUS; MEDICAGO SATIVA; Microbiology; Molecular biology; MOLECULAR SEQUENCE DATA; Mutation; NUCLEOTIDE SEQUENCE; Physics, chemistry, biochemistry and biology of agricultural and forest soils; Plant Microbiology; PUTA LOCUS; RHIZOBIACEAE; SECUENCIA NUCLEOTIDICA; SEQUENCE NUCLEOTIDIQUE; Sinorhizobium meliloti; Soil science; Soils; Root; Medicago sativa; Nodulation; Vegetal microorganism relation; Rhizosphere; Leguminosae; Gene; Betaine; Dicotyledones; Angiospermae; Bacteria; Spermatophyta; Catabolism; Rhizobiaceae; Mutation; Fodder crop; Colonization
• ISSN: 0099-2240; 1098-5336
• DOI: 10.1128/aem.64.10.3954-3960.1998

Stachydrine, a betaine released by germinating alfalfa seeds, functions as an inducer of nodulation genes, a catabolite, and an osmoprotectant in Sinorhizobium meliloti. Two stachydrine-inducible genes were found in S. meliloti 1021 by mutation with a Tn5-luxAB promoter probe. Both mutant strains (S10 and S11) formed effective alfalfa root nodules, but neither grew on stachydrine as the sole carbon and nitrogen source. When grown in the absence or presence of salt stress, S10 and S11 took up [14C]stachydrine as well as wild-type cells did, but neither used stachydrine effectively as an osmoprotectant. In the absence of salt stress, both S10 and S11 took up less [14C]proline than wild-type cells did. S10 and S11 appeared to colonize alfalfa roots normally in single-strain tests, but when mixed with the wild-type strain, their rhizosphere counts were reduced more than 50% (P less than or equal to 0.01) relative to the wild type. These results suggest that stachydrine catabolism contributes to root colonization. DNA sequence analysis identified the mutated locus in S11 as putA, and the luxAB fusion in that gene was induced by proline as well as stachydrine. DNA that restored the capacity of mutant S10 to catabolize stachydrine contained a new open reading frame, stcD. All data are consistent with the concept that stcD codes for an enzyme that produces proline by demethylation of N-methylproline, a degradation product of stachydrine