Inositol Catabolism, a Key Pathway in Sinorhizobium meliloti for Competitive Host Nodulation

ABSTRACT The nitrogen-fixing symbiont of alfalfa, Sinorhizobium meliloti, is able to use myo-inositol as the sole carbon source. Putative inositol catabolism genes (iolA and iolRCDEB) have been identified in the S. meliloti genome based on their similarities with the Bacillus subtilis iol genes. In this study, functional mutational analysis revealed that the iolA and iolCDEB genes are required for growth not only with the myo-isomer but also for growth with scyllo- and d-chiro-inositol as the sole carbon source. An additional, hypothetical dehydrogenase of the IdhA/MocA/GFO family encoded by the smc01163 gene was found to be essential for growth with scyllo-inositol, whereas the idhA-encoded myo-inositol dehydrogenase was responsible for the oxidation of d-chiro-inositol. The putative regulatory iolR gene, located upstream of iolCDEB, encodes a repressor of the iol genes, negatively regulating the activity of the myo- and the scyllo-inositol dehydrogenases. Mutants with insertions in the iolA, smc01163, and individual iolRCDE genes could not compete against the wild type in a nodule occupancy assay on alfalfa plants. Thus, a functional inositol catabolic pathway and its proper regulation are important nutritional or signaling factors in the S. meliloti-alfalfa symbiosis.

[1]  H. Ashida,et al.  Identification of two scyllo-inositol dehydrogenases in Bacillus subtilis. , 2010, Microbiology.

[2]  Serafim Batzoglou,et al.  Genetic and Computational Identification of a Conserved Bacterial Metabolic Module , 2008, PLoS genetics.

[3]  T. Fuchs,et al.  Characterization of the myo-Inositol Utilization Island of Salmonella enterica serovar Typhimurium † , 2008 .

[4]  Petra R. A. Kohler,et al.  Chemical synthesis of scyllo-inosamine and catabolism studies in Sinorhizobium meliloti. , 2008, Bioorganic & medicinal chemistry.

[5]  Y. Fujita,et al.  myo-Inositol Catabolism in Bacillus subtilis* , 2008, Journal of Biological Chemistry.

[6]  D. R. Palmer,et al.  Probing the promiscuous active site of myo-inositol dehydrogenase using synthetic substrates, homology modeling, and active site modification. , 2007, Biochemistry.

[7]  V. Monedero,et al.  Identification of a Gene Cluster Enabling Lactobacillus casei BL23 To Utilize myo-Inositol , 2007, Applied and Environmental Microbiology.

[8]  J. McLaurin,et al.  Properties of scyllo–inositol as a therapeutic treatment of AD-like pathology , 2007, Journal of Molecular Medicine.

[9]  Folker Meyer,et al.  Construction of a Large Signature-Tagged Mini-Tn5 Transposon Library and Its Application to Mutagenesis of Sinorhizobium meliloti , 2006, Applied and Environmental Microbiology.

[10]  H. Ashida,et al.  Genetic Modification of Bacillus subtilis for Production of d-chiro-Inositol, an Investigational Drug Candidate for Treatment of Type 2 Diabetes and Polycystic Ovary Syndrome , 2006, Applied and Environmental Microbiology.

[11]  A. Becker,et al.  Global transcriptional analysis of the phosphate starvation response in Sinorhizobium meliloti strains 1021 and 2011 , 2004, Molecular Genetics and Genomics.

[12]  Kayo Okumura,et al.  Organization and transcriptional regulation of myo-inositol operon in Clostridium perfringens. , 2004, FEMS microbiology letters.

[13]  H. Matsui,et al.  Structure and expression of 12-oxophytodienoate reductase (subgroup I) genes in pea, and characterization of the oxidoreductase activities of their recombinant products , 2004, Molecular Genetics and Genomics.

[14]  S. Long,et al.  Analysis of differences between Sinorhizobium meliloti 1021 and 2011 strains using the host calcium spiking response. , 2002, Molecular plant-microbe interactions : MPMI.

[15]  Benjamin L Turner,et al.  Inositol phosphates in the environment. , 2002, Philosophical transactions of the Royal Society of London. Series B, Biological sciences.

[16]  M. Wood,et al.  Investigation of myo-inositol catabolism in Rhizobium leguminosarum bv. viciae and its effect on nodulation competitiveness. , 2001, Molecular plant-microbe interactions : MPMI.

[17]  A. H. Krishnan,et al.  A Functional myo-Inositol Dehydrogenase Gene Is Required for Efficient Nitrogen Fixation and Competitiveness of Sinorhizobium fredii USDA191 To Nodulate Soybean (Glycine max [L.] Merr.) , 2001, Journal of bacteriology.

[18]  M. Wood,et al.  Myo-inositol catabolism by Rhizobium in soil: HPLC and enzymatic studies , 2001 .

[19]  K. Kobayashi,et al.  Combined transcriptome and proteome analysis as a powerful approach to study genes under glucose repression in Bacillus subtilis. , 2001, Nucleic acids research.

[20]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[21]  F. Loewus,et al.  myo-Inositol metabolism in plants , 2000 .

[22]  S. Long,et al.  Bacterial genes induced within the nodule during the Rhizobium–legume symbiosis , 1999, Molecular microbiology.

[23]  P. Murphy,et al.  A model for the catabolism of rhizopine in Rhizobium leguminosarum involves a ferredoxin oxygenase complex and the inositol degradative pathway. , 1998, Molecular plant-microbe interactions : MPMI.

[24]  Szi Fei Feng,et al.  A functional myo-inositol catabolism pathway is essential for rhizopine utilization by Sinorhizobium meliloti. , 1998, Microbiology.

[25]  Y. Fujita,et al.  Organization and transcription of the myo-inositol operon, iol, of Bacillus subtilis , 1997, Journal of Bacteriology.

[26]  F. D. de Bruijn,et al.  Structural and functional conservation of the rhizopine catabolism (moc) locus is limited to selected Rhizobium meliloti strains and unrelated to their geographical origin. , 1995, Molecular plant-microbe interactions : MPMI.

[27]  P. Murphy,et al.  The distribution of inositol rhizopine genes in Rhizobium populations , 1995 .

[28]  Frans J. de Bruijn,et al.  Molecular and genetic characterization of the rhizopine catabolism (mocABRC) genes of Rhizobium meliloti L5-30 , 1994, Molecular and General Genetics MGG.

[29]  P. Poole,et al.  myo-lnositol catabolism and catabolite regulation in Rhizobium leguminosarum bv . viciae , 2022 .

[30]  Y. Fujita,et al.  Cloning and nucleotide sequencing of a 15 kb region of the Bacillus subtilis genome containing the iol operon. , 1994, Microbiology.

[31]  M. Hynes,et al.  Versatile suicide vectors which allow direct selection for gene replacement in gram-negative bacteria. , 1993, Gene.

[32]  James A. Duke,et al.  Handbook of Phytochemical Constituents of GRAS Herbs and Other Economic Plants , 1992 .

[33]  J. Streeter.  Carbohydrate, organic Acid, and amino Acid composition of bacteroids and cytosol from soybean nodules. , 1987, Plant physiology.

[34]  T. Finan,et al.  Second symbiotic megaplasmid in Rhizobium meliloti carrying exopolysaccharide and thiamine synthesis genes , 1986, Journal of bacteriology.

[35]  T. Egelhoff,et al.  Rhizobium meliloti nodulation genes: identification of nodDABC gene products, purification of nodA protein, and expression of nodA in Rhizobium meliloti , 1985, Journal of bacteriology.

[36]  H. Krisch,et al.  In vitro insertional mutagenesis with a selectable DNA fragment. , 1984, Gene.

[37]  Susan E. Brown,et al.  Physical and genetic characterization of symbiotic and auxotrophic mutants of Rhizobium meliloti induced by transposon Tn5 mutagenesis , 1982, Journal of bacteriology.

[38]  R. Ramaley,et al.  Purification and properties of Bacillus subtilis inositol dehydrogenase. , 1979, The Journal of biological chemistry.

[39]  E. Vincze,et al.  Genetic and Biochemical Analysis of Mutants Affected in Nitrate Reduction in Rhizobium meliloti , 1979 .

[40]  D. Helinski,et al.  Replication of an origin-containing derivative of plasmid RK2 dependent on a plasmid function provided in trans. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[41]  H. Meade,et al.  Genetic mapping of Rhizobium meliloti. , 1977, Proceedings of the National Academy of Sciences of the United States of America.

[42]  J. Beringer.  R factor transfer in Rhizobium leguminosarum. , 1974, Journal of general microbiology.

[43]  W. Broughton,et al.  Control of leghaemoglobin synthesis in snake beans. , 1971, The Biochemical journal.

[44]  B. Magasanik,et al.  The pathway of myo-inositol degradation in Aerobacter aerogenes. Dehydrogenation and dehydration. , 1966, The Journal of biological chemistry.

[45]  Herbert E. Carter,et al.  Oxidation of inositol by Acetobacter suboxydans. , 1948, The Journal of biological chemistry.

[46]  R. A. M. Departrneri.  Myo-inositol Catabolism in Salmonella typhimurium : Enzyme Repression Dependent on Growth History of Organism , 2008 .

[47]  T. Beveridge,et al.  Methods for general and molecular microbiology , 2007 .

[48]  J. Larner,et al.  D-Chiro-Inositol – Its Functional Role in Insulin Action and its Deficit in Insulin Resistance , 2002, International journal of experimental diabetes research.

[49]  L. L. Voelker,et al.  Transposon mutagenesis. , 1998, Methods in molecular biology.

[50]  R. Jefferson,et al.  The Escherichia coli gus operon: induction and expression of the gus operon in E. coli and the occurrence and use of GUS in other bacteria. , 1992 .

[51]  Z. Bánfalvi,et al.  Genes for the catabolism and synthesis of an opine-like compound in Rhizobium meliloti are closely linked and on the Sym plasmid. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[52]  L. Anderson.  15 – THE CYCLITOLS , 1972 .