Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1996 Aug;178(15):4635–4642. doi: 10.1128/jb.178.15.4635-4642.1996

Beta-glucan synthesis in Bradyrhizobium japonicum: characterization of a new locus (ndvC) influencing beta-(1-->6) linkages.

A A Bhagwat 1, K C Gross 1, R E Tully 1, D L Keister 1
PMCID: PMC178234  PMID: 8755895

Abstract

Bradyrhizobium japonicum synthesizes periplasmic cyclic beta-(1-->3),beta-(1-->6)-D-glucans during growth in hypoosmotic environments, and evidence is growing that these molecules may have a specific function during plant-microbe interactions in addition to osmoregulation. Site-directed Tn5 mutagenesis of the DNA region upstream of ndvB resulted in identification of a new gene (ndvC) involved in beta-(1--> 3), beta-(1-->6)-glucan synthesis and in nodule development. The predicted translation product was a polypeptide (ca. 62 kDa) with several transmembrane domains. It contained a sequence characteristic of a conserved nucleoside-sugar-binding motif found in many bacterial enzymes and had 51% similarity with a beta-glucanosyltransferase from Candida albicans. B. japonicum carrying a Tn5 insertion in ndvC resulted in synthesis of altered cyclic beta-glucans composed almost entirely of beta-(1--> 3)-glycosyl linkages. The mutant strain was only slightly sensitive to hypoosmotic growth conditions compared with the ndvB mutant, but it was severely impaired in symbiotic interactions with soybean (Glycine max). Nodulation was delayed by 8 to 10 days, and many small nodule-like structures apparently devoid of viable bacteria were formed. This finding suggests that the structure of the beta-glucan molecule is important for a successful symbiotic interaction, and beta-glucans may have a specific function in addition to their role in hypoosmotic adaptation.

Full Text

The Full Text of this article is available as a PDF (717.1 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. Basic local alignment search tool. J Mol Biol. 1990 Oct 5;215(3):403–410. doi: 10.1016/S0022-2836(05)80360-2. [DOI] [PubMed] [Google Scholar]
  2. Basse C. W., Bock K., Boller T. Elicitors and suppressors of the defense response in tomato cells. Purification and characterization of glycopeptide elicitors and glycan suppressors generated by enzymatic cleavage of yeast invertase. J Biol Chem. 1992 May 25;267(15):10258–10265. [PubMed] [Google Scholar]
  3. Bhagwat A. A., Tully R. E., Keister D. L. Identification and cloning of a cyclic beta-(1-->3), beta-(1-->6)-D-glucan synthesis locus from Bradyrhizobium japonicum. FEMS Microbiol Lett. 1993 Dec 1;114(2):139–144. doi: 10.1111/j.1574-6968.1993.tb06564.x. [DOI] [PubMed] [Google Scholar]
  4. Bhagwat A. A., Tully R. E., Keister D. L. Isolation and characterization of an ndvB locus from Rhizobium fredii. Mol Microbiol. 1992 Aug;6(15):2159–2165. doi: 10.1111/j.1365-2958.1992.tb01389.x. [DOI] [PubMed] [Google Scholar]
  5. Breedveld M. W., Miller K. J. Cyclic beta-glucans of members of the family Rhizobiaceae. Microbiol Rev. 1994 Jun;58(2):145–161. doi: 10.1128/mr.58.2.145-161.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cangelosi G. A., Hung L., Puvanesarajah V., Stacey G., Ozga D. A., Leigh J. A., Nester E. W. Common loci for Agrobacterium tumefaciens and Rhizobium meliloti exopolysaccharide synthesis and their roles in plant interactions. J Bacteriol. 1987 May;169(5):2086–2091. doi: 10.1128/jb.169.5.2086-2091.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cangelosi G. A., Martinetti G., Leigh J. A., Lee C. C., Thienes C., Theines C., Nester E. W. Role for [corrected] Agrobacterium tumefaciens ChvA protein in export of beta-1,2-glucan. J Bacteriol. 1989 Mar;171(3):1609–1615. doi: 10.1128/jb.171.3.1609-1615.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cangelosi G. A., Martinetti G., Nester E. W. Osmosensitivity phenotypes of Agrobacterium tumefaciens mutants that lack periplasmic beta-1,2-glucan. J Bacteriol. 1990 Apr;172(4):2172–2174. doi: 10.1128/jb.172.4.2172-2174.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Clarke H. R., Leigh J. A., Douglas C. J. Molecular signals in the interactions between plants and microbes. Cell. 1992 Oct 16;71(2):191–199. doi: 10.1016/0092-8674(92)90348-g. [DOI] [PubMed] [Google Scholar]
  10. Cohen J. L., Miller K. J. A novel membrane-bound glucosyltransferase from Bradyrhizobium japonicum. J Bacteriol. 1991 Jul;173(14):4271–4276. doi: 10.1128/jb.173.14.4271-4276.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Cole M. A., Elkan G. H. Transmissible resistance to penicillin G, neomycin, and chloramphenicol in Rhizobium japonicum. Antimicrob Agents Chemother. 1973 Sep;4(3):248–253. doi: 10.1128/aac.4.3.248. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Cosio E. G., Frey T., Ebel J. Identification of a high-affinity binding protein for a hepta-beta-glucoside phytoalexin elicitor in soybean. Eur J Biochem. 1992 Mar 15;204(3):1115–1123. doi: 10.1111/j.1432-1033.1992.tb16736.x. [DOI] [PubMed] [Google Scholar]
  13. Dylan T., Helinski D. R., Ditta G. S. Hypoosmotic adaptation in Rhizobium meliloti requires beta-(1----2)-glucan. J Bacteriol. 1990 Mar;172(3):1400–1408. doi: 10.1128/jb.172.3.1400-1408.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Dylan T., Ielpi L., Stanfield S., Kashyap L., Douglas C., Yanofsky M., Nester E., Helinski D. R., Ditta G. Rhizobium meliloti genes required for nodule development are related to chromosomal virulence genes in Agrobacterium tumefaciens. Proc Natl Acad Sci U S A. 1986 Jun;83(12):4403–4407. doi: 10.1073/pnas.83.12.4403. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Farkas I., Hardy T. A., DePaoli-Roach A. A., Roach P. J. Isolation of the GSY1 gene encoding yeast glycogen synthase and evidence for the existence of a second gene. J Biol Chem. 1990 Dec 5;265(34):20879–20886. [PubMed] [Google Scholar]
  16. Fath M. J., Kolter R. ABC transporters: bacterial exporters. Microbiol Rev. 1993 Dec;57(4):995–1017. doi: 10.1128/mr.57.4.995-1017.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Fu C., Maier R. J. Rapid and efficient selection of recombinant site-directed mutants of Bradyrhizobium japonicum by colony hybridization. FEMS Microbiol Lett. 1993 May 1;109(1):33–38. doi: 10.1111/j.1574-6968.1993.tb06139.x. [DOI] [PubMed] [Google Scholar]
  18. Furukawa K., Tagaya M., Inouye M., Preiss J., Fukui T. Identification of lysine 15 at the active site in Escherichia coli glycogen synthase. Conservation of Lys-X-Gly-Gly sequence in the bacterial and mammalian enzymes. J Biol Chem. 1990 Feb 5;265(4):2086–2090. [PubMed] [Google Scholar]
  19. Gore R. S., Miller K. J. Cyclic [beta]-1,6 -1,3 Glucans Are Synthesized by Bradyrhizobium japonicum Bacteroids within Soybean (Glycine max) Root Nodules. Plant Physiol. 1993 May;102(1):191–194. doi: 10.1104/pp.102.1.191. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hartland R. P., Emerson G. W., Sullivan P. A. A secreted beta-glucan-branching enzyme from Candida albicans. Proc Biol Sci. 1991 Nov 22;246(1316):155–160. doi: 10.1098/rspb.1991.0138. [DOI] [PubMed] [Google Scholar]
  21. Høj P. B., Condron R., Traeger J. C., McAuliffe J. C., Stone B. A. Identification of glutamic acid 105 at the active site of Bacillus amyloliquefaciens 1,3-1,4-beta-D-glucan 4-glucanohydrolase using epoxide-based inhibitors. J Biol Chem. 1992 Dec 15;267(35):25059–25066. [PubMed] [Google Scholar]
  22. Ielpi L., Dylan T., Ditta G. S., Helinski D. R., Stanfield S. W. The ndvB locus of Rhizobium meliloti encodes a 319-kDa protein involved in the production of beta-(1----2)-glucan. J Biol Chem. 1990 Feb 15;265(5):2843–2851. [PubMed] [Google Scholar]
  23. Klebl F., Tanner W. Molecular cloning of a cell wall exo-beta-1,3-glucanase from Saccharomyces cerevisiae. J Bacteriol. 1989 Nov;171(11):6259–6264. doi: 10.1128/jb.171.11.6259-6264.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Klis F. M. Review: cell wall assembly in yeast. Yeast. 1994 Jul;10(7):851–869. doi: 10.1002/yea.320100702. [DOI] [PubMed] [Google Scholar]
  25. Kyte J., Doolittle R. F. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982 May 5;157(1):105–132. doi: 10.1016/0022-2836(82)90515-0. [DOI] [PubMed] [Google Scholar]
  26. Leigh J. A., Coplin D. L. Exopolysaccharides in plant-bacterial interactions. Annu Rev Microbiol. 1992;46:307–346. doi: 10.1146/annurev.mi.46.100192.001515. [DOI] [PubMed] [Google Scholar]
  27. Linthorst H. J., Melchers L. S., Mayer A., van Roekel J. S., Cornelissen B. J., Bol J. F. Analysis of gene families encoding acidic and basic beta-1,3-glucanases of tobacco. Proc Natl Acad Sci U S A. 1990 Nov;87(22):8756–8760. doi: 10.1073/pnas.87.22.8756. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Long S. R., Staskawicz B. J. Prokaryotic plant parasites. Cell. 1993 Jun 4;73(5):921–935. doi: 10.1016/0092-8674(93)90271-q. [DOI] [PubMed] [Google Scholar]
  29. Miller K. J., Gore R. S., Johnson R., Benesi A. J., Reinhold V. N. Cell-associated oligosaccharides of Bradyrhizobium spp. J Bacteriol. 1990 Jan;172(1):136–142. doi: 10.1128/jb.172.1.136-142.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Miller K. J., Hadley J. A., Gustine D. L. Cyclic [beta]-1,6-1,3-Glucans of Bradyrhizobium japonicum USDA 110 Elicit Isoflavonoid Production in the Soybean (Glycine max) Host. Plant Physiol. 1994 Mar;104(3):917–923. doi: 10.1104/pp.104.3.917. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Miller K. J., Kennedy E. P., Reinhold V. N. Osmotic adaptation by gram-negative bacteria: possible role for periplasmic oligosaccharides. Science. 1986 Jan 3;231(4733):48–51. doi: 10.1126/science.3941890. [DOI] [PubMed] [Google Scholar]
  32. Mrsa V., Klebl F., Tanner W. Purification and characterization of the Saccharomyces cerevisiae BGL2 gene product, a cell wall endo-beta-1,3-glucanase. J Bacteriol. 1993 Apr;175(7):2102–2106. doi: 10.1128/jb.175.7.2102-2106.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. O'Connell K. P., Handelsman J. chvA locus may be involved in export of neutral cyclic beta-1,2-linked D-glucan from Agrobacterium tumefaciens. Mol Plant Microbe Interact. 1989 Jan-Feb;2(1):11–16. [PubMed] [Google Scholar]
  34. Pfeffer P. E., Bécard G., Rolin D. B., Uknalis J., Cooke P., Tu S. In vivo nuclear magnetic resonance study of the osmoregulation of phosphocholine-substituted beta-1,3;1,6 cyclic glucan and its associated carbon metabolism in Bradyrhizobium japonicum USDA 110. Appl Environ Microbiol. 1994 Jun;60(6):2137–2146. doi: 10.1128/aem.60.6.2137-2146.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Roemer T., Paravicini G., Payton M. A., Bussey H. Characterization of the yeast (1-->6)-beta-glucan biosynthetic components, Kre6p and Skn1p, and genetic interactions between the PKC1 pathway and extracellular matrix assembly. J Cell Biol. 1994 Oct;127(2):567–579. doi: 10.1083/jcb.127.2.567. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Rolin D. B., Pfeffer P. E., Osman S. F., Szwergold B. S., Kappler F., Benesi A. J. Structural studies of a phosphocholine substituted beta-(1,3);(1,6) macrocyclic glucan from Bradyrhizobium japonicum USDA 110. Biochim Biophys Acta. 1992 Jun 12;1116(3):215–225. doi: 10.1016/0304-4165(92)90014-l. [DOI] [PubMed] [Google Scholar]
  37. Seki N., Muta T., Oda T., Iwaki D., Kuma K., Miyata T., Iwanaga S. Horseshoe crab (1,3)-beta-D-glucan-sensitive coagulation factor G. A serine protease zymogen heterodimer with similarities to beta-glucan-binding proteins. J Biol Chem. 1994 Jan 14;269(2):1370–1374. [PubMed] [Google Scholar]
  38. Stanfield S. W., Ielpi L., O'Brochta D., Helinski D. R., Ditta G. S. The ndvA gene product of Rhizobium meliloti is required for beta-(1----2)glucan production and has homology to the ATP-binding export protein HlyB. J Bacteriol. 1988 Aug;170(8):3523–3530. doi: 10.1128/jb.170.8.3523-3530.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Tully R. E., Keister D. L., Gross K. C. Fractionation of the beta-Linked Glucans of Bradyrhizobium japonicum and Their Response to Osmotic Potential. Appl Environ Microbiol. 1990 Jun;56(6):1518–1522. doi: 10.1128/aem.56.6.1518-1522.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. York W. S., McNeil M., Darvill A. G., Albersheim P. Beta-2-linked glucans secreted by fast-growing species of Rhizobium. J Bacteriol. 1980 Apr;142(1):243–248. doi: 10.1128/jb.142.1.243-248.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Zorreguieta A., Cavaignac S., Geremia R. A., Ugalde R. A. Osmotic regulation of beta(1-2) glucan synthesis in members of the family Rhizobiaceae. J Bacteriol. 1990 Aug;172(8):4701–4704. doi: 10.1128/jb.172.8.4701-4704.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Zorreguieta A., Ugalde R. A. Formation in Rhizobium and Agrobacterium spp. of a 235-kilodalton protein intermediate in beta-D(1-2) glucan synthesis. J Bacteriol. 1986 Sep;167(3):947–951. doi: 10.1128/jb.167.3.947-951.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES