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. 1991 May;173(9):2809–2817. doi: 10.1128/jb.173.9.2809-2817.1991

Genetic analysis of a region of the Rhizobium meliloti pSym plasmid specifying catabolism of trigonelline, a secondary metabolite present in legumes.

C Boivin 1, L R Barran 1, C A Malpica 1, C Rosenberg 1
PMCID: PMC207861  PMID: 1850402

Abstract

Genes controlling the catabolism of trigonelline, a secondary metabolite that is often present in legumes, are located on the pSym megaplasmid of Rhizobium meliloti. To investigate the role of bacterial trigonelline catabolism in the Rhizobium-legume symbiosis, we identified and characterized the R. meliloti RCR2011 genetic loci (trc) controlling trigonelline catabolism. Tn5-B20 mutagenesis showed that the trc region is a continuous DNA segment of 9 kb located 4 kb downstream of the nifAB and fdxN genes. Trc mutants fell into two classes according to their phenotype and location: (i) mutants carrying Tn5-B20 insertions in the right-hand part of the trc region were incapable of growing on trigonelline as the sole carbon and/or nitrogen source, and (ii) insertions in the left-hand part of the trc region resulted in delayed growth on trigonelline as the sole carbon and/or nitrogen source. No significant defect in nodule formation or nitrogen fixation was detected for mutants of either class. Screening of a set of R. meliloti strains from various geographical origins showed that all of these strains are able to catabolize trigonelline and show sequence homology between their megaplasmids and a trc probe.

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Selected References

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  1. Banfalvi Z., Kondorosi E., Kondorosi A. Rhizobium meliloti carries two megaplasmids. Plasmid. 1985 Mar;13(2):129–138. doi: 10.1016/0147-619x(85)90065-4. [DOI] [PubMed] [Google Scholar]
  2. Boivin C., Camut S., Malpica C. A., Truchet G., Rosenberg C. Rhizobium meliloti Genes Encoding Catabolism of Trigonelline Are Induced under Symbiotic Conditions. Plant Cell. 1990 Dec;2(12):1157–1170. doi: 10.1105/tpc.2.12.1157. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Boucher C. A., Van Gijsegem F., Barberis P. A., Arlat M., Zischek C. Pseudomonas solanacearum genes controlling both pathogenicity on tomato and hypersensitivity on tobacco are clustered. J Bacteriol. 1987 Dec;169(12):5626–5632. doi: 10.1128/jb.169.12.5626-5632.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bánfalvi Z., Sakanyan V., Koncz C., Kiss A., Dusha I., Kondorosi A. Location of nodulation and nitrogen fixation genes on a high molecular weight plasmid of R. meliloti. Mol Gen Genet. 1981;184(2):318–325. doi: 10.1007/BF00272925. [DOI] [PubMed] [Google Scholar]
  5. De Vos G. F., Walker G. C., Signer E. R. Genetic manipulations in Rhizobium meliloti utilizing two new transposon Tn5 derivatives. Mol Gen Genet. 1986 Sep;204(3):485–491. doi: 10.1007/BF00331029. [DOI] [PubMed] [Google Scholar]
  6. Debellé F., Rosenberg C., Vasse J., Maillet F., Martinez E., Dénarié J., Truchet G. Assignment of symbiotic developmental phenotypes to common and specific nodulation (nod) genetic loci of Rhizobium meliloti. J Bacteriol. 1986 Dec;168(3):1075–1086. doi: 10.1128/jb.168.3.1075-1086.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  8. Finan T. M., Kunkel B., De Vos G. F., Signer E. R. Second symbiotic megaplasmid in Rhizobium meliloti carrying exopolysaccharide and thiamine synthesis genes. J Bacteriol. 1986 Jul;167(1):66–72. doi: 10.1128/jb.167.1.66-72.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Friedman A. M., Long S. R., Brown S. E., Buikema W. J., Ausubel F. M. Construction of a broad host range cosmid cloning vector and its use in the genetic analysis of Rhizobium mutants. Gene. 1982 Jun;18(3):289–296. doi: 10.1016/0378-1119(82)90167-6. [DOI] [PubMed] [Google Scholar]
  10. Hardy R. W., Holsten R. D., Jackson E. K., Burns R. C. The acetylene-ethylene assay for n(2) fixation: laboratory and field evaluation. Plant Physiol. 1968 Aug;43(8):1185–1207. doi: 10.1104/pp.43.8.1185. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Klipp W., Reiländer H., Schlüter A., Krey R., Pühler A. The Rhizobium meliloti fdxN gene encoding a ferredoxin-like protein is necessary for nitrogen fixation and is cotranscribed with nifA and nifB. Mol Gen Genet. 1989 Apr;216(2-3):293–302. doi: 10.1007/BF00334368. [DOI] [PubMed] [Google Scholar]
  12. Kuo Y. H., Lambein F., Ikegami F., Van Parijs R. Isoxazolin-5-ones and Amino Acids in Root Exudates of Pea and Sweet Pea Seedlings. Plant Physiol. 1982 Nov;70(5):1283–1289. doi: 10.1104/pp.70.5.1283. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Le Rudulier D., Bouillard L. Glycine betaine, an osmotic effector in Klebsiella pneumoniae and other members of the Enterobacteriaceae. Appl Environ Microbiol. 1983 Jul;46(1):152–159. doi: 10.1128/aem.46.1.152-159.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Long S. R. Rhizobium-legume nodulation: life together in the underground. Cell. 1989 Jan 27;56(2):203–214. doi: 10.1016/0092-8674(89)90893-3. [DOI] [PubMed] [Google Scholar]
  15. Martin M. O., Long S. R. Generalized transduction in Rhizobium meliloti. J Bacteriol. 1984 Jul;159(1):125–129. doi: 10.1128/jb.159.1.125-129.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Melchers L. S., Regensburg-Tuïnk A. J., Schilperoort R. A., Hooykaas P. J. Specificity of signal molecules in the activation of Agrobacterium virulence gene expression. Mol Microbiol. 1989 Jul;3(7):969–977. doi: 10.1111/j.1365-2958.1989.tb00246.x. [DOI] [PubMed] [Google Scholar]
  17. Murphy P. J., Heycke N., Trenz S. P., Ratet P., de Bruijn F. J., Schell J. Synthesis of an opine-like compound, a rhizopine, in alfalfa nodules is symbiotically regulated. Proc Natl Acad Sci U S A. 1988 Dec;85(23):9133–9137. doi: 10.1073/pnas.85.23.9133. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Renalier M. H., Batut J., Ghai J., Terzaghi B., Gherardi M., David M., Garnerone A. M., Vasse J., Truchet G., Huguet T. A new symbiotic cluster on the pSym megaplasmid of Rhizobium meliloti 2011 carries a functional fix gene repeat and a nod locus. J Bacteriol. 1987 May;169(5):2231–2238. doi: 10.1128/jb.169.5.2231-2238.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Rolfe B. G. Flavones and isoflavones as inducing substances of legume nodulation. Biofactors. 1988 Jan;1(1):3–10. [PubMed] [Google Scholar]
  20. Rosenberg C., Boistard P., Dénarié J., Casse-Delbart F. Genes controlling early and late functions in symbiosis are located on a megaplasmid in Rhizobium meliloti. Mol Gen Genet. 1981;184(2):326–333. doi: 10.1007/BF00272926. [DOI] [PubMed] [Google Scholar]
  21. Scott D. B., Wilson R., Shaw G. J., Petit A., Tempe J. Biosynthesis and degradation of nodule-specific Rhizobium loti compounds in Lotus nodules. J Bacteriol. 1987 Jan;169(1):278–282. doi: 10.1128/jb.169.1.278-282.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Simon R., Quandt J., Klipp W. New derivatives of transposon Tn5 suitable for mobilization of replicons, generation of operon fusions and induction of genes in gram-negative bacteria. Gene. 1989 Aug 1;80(1):161–169. doi: 10.1016/0378-1119(89)90262-x. [DOI] [PubMed] [Google Scholar]
  23. Smith G. E., Summers M. D. The bidirectional transfer of DNA and RNA to nitrocellulose or diazobenzyloxymethyl-paper. Anal Biochem. 1980 Nov 15;109(1):123–129. doi: 10.1016/0003-2697(80)90019-6. [DOI] [PubMed] [Google Scholar]
  24. Smith L. T., Pocard J. A., Bernard T., Le Rudulier D. Osmotic control of glycine betaine biosynthesis and degradation in Rhizobium meliloti. J Bacteriol. 1988 Jul;170(7):3142–3149. doi: 10.1128/jb.170.7.3142-3149.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Stirling D. A., Hulton C. S., Waddell L., Park S. F., Stewart G. S., Booth I. R., Higgins C. F. Molecular characterization of the proU loci of Salmonella typhimurium and Escherichia coli encoding osmoregulated glycine betaine transport systems. Mol Microbiol. 1989 Aug;3(8):1025–1038. doi: 10.1111/j.1365-2958.1989.tb00253.x. [DOI] [PubMed] [Google Scholar]
  26. Tepfer D., Goldmann A., Pamboukdjian N., Maille M., Lepingle A., Chevalier D., Dénarié J., Rosenberg C. A plasmid of Rhizobium meliloti 41 encodes catabolism of two compounds from root exudate of Calystegium sepium. J Bacteriol. 1988 Mar;170(3):1153–1161. doi: 10.1128/jb.170.3.1153-1161.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Thacker R., Rørvig O., Kahlon P., Gunsalus I. C. NIC, a conjugative nicotine-nicotinate degradative plasmid in Pseudomonas convexa. J Bacteriol. 1978 Jul;135(1):289–290. doi: 10.1128/jb.135.1.289-290.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Truchet G., Debellé F., Vasse J., Terzaghi B., Garnerone A. M., Rosenberg C., Batut J., Maillet F., Dénarié J. Identification of a Rhizobium meliloti pSym2011 region controlling the host specificity of root hair curling and nodulation. J Bacteriol. 1985 Dec;164(3):1200–1210. doi: 10.1128/jb.164.3.1200-1210.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]

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