Skip to main content
SpringerLink
Log in

The formation of R-prime deletion mutants and the identification of the symbiotic genes in Rhizobium fredii strain USDA191

  • Published:
Plant Molecular Biology Aims and scope Submit manuscript

Summary

R-prime plasmids were formed between the plasmid of Rhizobium fredii strain USDA191 containing nodulation and nitrogen-fixation genes, pRjaUSDA191c, and pRL180, and RP1 derivative. R. fredii USDA191 contains four HindIII fragments that hybridize with an 8.7 kb EcoRI fragment that contains nodulation genes from R. meliloti. These four fragments are on pRjaUSDA191c and are 15.5 kb, 12.5 kb, 6.8 kb, and 5.2 kb in size. A series of R-primes generated in E. coli of pRjaUSDA191c were transferred into a Nod- Nif- derivative of strain USDA191 to determine which nodulation region is necessary for nodule formation. Transconjugants containing the 12.5 kb and the 6.8 kb HindIII fragments on segments of pRjaUSDA191c produced nodules on soybean plants. However, transconjugants containing the 12.5 kb HindIII fragment alone were unable to form nodules, suggesting that the 6.8 kb HindIII fragment or the 6.8 kb and the 12.5 kb HindIII fragments together were needed for nodule formation. The 6.8 kb HindIII fragment was subcloned into the vector pVK102 and transferred into transconjugants containing no sequences homologous to R. meliloti nodulation DNA or to transconjugants containing only the 12.5 kb HindIII fragment. Nodules were formed on soybeans only when both the 12.5 kb and the 6.8 kb HindIII fragments were present in R. frediistrain USDA191.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Banfalvi Z, Sakanyan V, Konaz 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 184:318–325, 1981.

    Google Scholar 

  2. Banfalvi Z, Randhawa G, Kondorosi E, Kiss A, Kondorosi A: Construction and characterization of R-prime plasmids carrying symbiotic genes of R. meliloti. Mol Gen Genet 189:129–135, 1983.

    Google Scholar 

  3. Beyon JL, Beringer JE, Johnston AWB: Plasmids and host range in Rhizobium leguminosarum and Rhizobium phaseoli. J Gen Microbiol 120:421–429, 1980.

    Google Scholar 

  4. Beringer JE: R factor transfer in R. japonicum. J Gen Microbiol 84:188–198, 1974.

    Google Scholar 

  5. Burris RH: Nitrogen fixation-assay methods and techniques. Methods Enzymol 24:415–431, 1972.

    Google Scholar 

  6. Ditta G, Stanfield S, Corbin D, Helinski DR: Broad host range cloning system for gram-negative bacteria: Construction of a gene bank of Rhizobium meliloti. Proc Natl Acad Sci USA 77:7347–7351, 1980.

    Google Scholar 

  7. Eckhardt T: A rapid method for the identification of plasmid deoxyribonucleic acid in bacteria. Plasmid 1:584–588, 1978.

    Google Scholar 

  8. Enquist L, Sternberg N: In vitro packaging of λ Dam vectors and their use in cloning DNA fragments. Methods Enzymol 68:281–284, 1979.

    Google Scholar 

  9. Grunstein M, Hogness DS: Colony hybridization: A method for the isolation of cloned DNAs that contain a specific gene. Proc Natl Acad Sci USA 72:3961–3965, 1975.

    Google Scholar 

  10. Hattori J, Johnson DA: Fast-growing Rhizobium japonicum that effectively nodulates several commercial Glycine max L. Merrill cultivars. Appl Environ Microbiol 48:234–235, 1984.

    Google Scholar 

  11. Heron DS, Pueppke SG: Mode of infection, nodulation specificity, and indigenous plasmids of 11 fast-growing Rhizobium japonicum strains. J Bacteriol 160:1061–1066, 1984.

    Google Scholar 

  12. Hirsch AM, Wilson KJ, Jones JDG, Bang M, Walder VV, Ausubel FM: Rhizobium meliloti nodulation genes allow Agrobacterium tumefaciens and Escherichia coli to form pseudonodules on alfalfa. J Bacteriol 158:1133–1143, 1984.

    Google Scholar 

  13. Hirsch PR, Van Montagu M, Johnston AWB, Brewin NJ, Schell J: Physcial identification of bacteriocinogenic, nodulation and other plasmids in strains of Rhizobium leguminosarum. J Gen Microbiol 120:403–412, 1980.

    Google Scholar 

  14. Holmes DS, Quigley M: A rapid boiling method for the preparation of bacterial plasmids. Anal Biochem 114:193, 1981.

    Google Scholar 

  15. Hombrecher G, Brewin NJ, Johnston AWB: Linkage of genes for nitrogenase and nodulation ability on plasmids in Rhizobium leguminosarum and R. phaseoli. Mol Gen Genet 182:133–136, 1981.

    Google Scholar 

  16. Hooykaas PJJ, den Dulk-Ras H, Schilperoort RA: Method for the transfer of large cryptic, non-self-transmissible plasmids: Ex planta transfer of the virulence plasmid of Agrobacterium rhizogenes. Plasmid 8:94–96, 1982a.

    Google Scholar 

  17. Hooykaas PJJ, Snijdewint FGM, Schilperoort RA: Identification of the Sym plasmid of Rhizobium leguminosarum strain 1001 and its transfer to and expression in other Rhizobia and Agrobacterium tumefaciens. Plasmid 8:73–82, 1982b.

    Google Scholar 

  18. Hooykaas PJJ, van Brussel AAN, den Dulk-Ras H, van Slogteren GMS, Schilperoort RA: Sym plasmid of Rhizobium trifolii expressed in different rhizobial species and Agrobacterium tumefaciens. Nature (London) 291:351–353, 1981.

    Google Scholar 

  19. Jansenvan Rensburg H, Strijdom BW, Otto CJ: Effective nodulation of soybeans by fast-growing strains of Rhizobium japonicum. S Afr J Sci 79:251–252, 1983.

    Google Scholar 

  20. Johnston AWB, Benyon JL, Buchanan-Wollaston AV, Setchell SM, Hirsch PR, Beringer JE: High frequency transfer of nodulating ability between strains and species of Rhizobium. Nature (London) 276:635–636, 1978.

    Google Scholar 

  21. Julliot JS, Dusha I, Renalier MH, Terzaghi B, Garnerone AM, Boistard P: An RP4-prime containing a 285 kb fragment of Rhizobium meliloti pSym megaplasmid: structural characterization and utilization for genetic studies of symbiotic functions controlled by pSym. Mol Gen Genet 193:17–26, 1984.

    Google Scholar 

  22. Keyser HH, Bohlool BB, Hu TS, Weber DF: Fast-growing Rhizobia isolated from root nodules of soybean. Science 215:1631–1632, 1982.

    Google Scholar 

  23. Knauf V, Nester E: Wide host range cloning vectors: a cosmid clone bank of an Agrobacterium Ti plasmid. Plasmid 8:45–54, 1982.

    Google Scholar 

  24. Kondorosi A, Kondorosi E, Pankhurst CE, Broughton WJ, Banfalvi Z: Mobilization of a Rhizobium meliloti megaplasmid carrying nodulation and nitrogen fixation genes into other Rhizobia and Agrobacterium. Mol Gen Genet 188:433–439, 1982.

    Google Scholar 

  25. Kondorosi E, Banfalvi A, Kondorosi A: Physical and genetic analysis of a symbiotic region of Rhizobium meliloti: Identification of nodulation genes. Mol Gen Genet 193:445–452, 1984.

    Google Scholar 

  26. Lamb JW, Hombrecher G, Johnston AWB: Plasmid-determined nodulation and nitrogen fixation abilities in Rhizobium phaseoli. Mol Gen Genet 186:449–452, 1982.

    Google Scholar 

  27. Long SL, Buikema WJ, Ausubel FM: Cloning of Rhizobium meliloti nodulation genes by direct complementation of Nod - mutants. Nature (London) 298:485–488, 1982.

    Google Scholar 

  28. Malik NSA, Pence MK, Calvert HE, Bauer WD: Rhizobium infection and nodule development in soybean are affected by exposure of the cotyledons to light. Plant Physiol 75:90–94, 1984.

    Google Scholar 

  29. Maniatis T, Fritsch EF, Sambrook J: Molecular Cloning: A Laboratory Manual. Cold Spring Harbour Laboratory, New York, 1982, pp 382–389.

    Google Scholar 

  30. Masterson RV, Russell PR, Atherly AG: Nitrogen fixation (nif) genes and large plasmids of Rhizobium japonicum. J Bacteriol 152:928–931, 1982.

    Google Scholar 

  31. Morrison NA, Cen YH, Chen HC, Plazinski J, Ridge R, Rolfe BG: Mobilization of a sym plasmid from a fast-growing cowpea Rhizobium strain. J Bacteriol 160:483–487, 1984.

    Google Scholar 

  32. Nuti MP, Lepidi AA, Prakash RK, Schilperoort RA, Cannon FC: Evidence for nitrogen fixation (nif) genes on indigenous Rhizobium plasmids. Nature (London) 282:533–535, 1979.

    Google Scholar 

  33. Olson ER, Sadowsky MJ, Verma DPS: Identification of genes involved in the Rhizobium-legume symbiosis by mudI (Kan, lac)-generated transcription fusions. Biotechnology 2:143–149, 1985.

    Google Scholar 

  34. Prakash RK, Atherly AG: Reiteration of genes involved in symbiotic nitrogen fixation by fast-growing Rhizobium japonicum. J Bacteriol 160:785–787, 1984.

    Google Scholar 

  35. Prakash RK, Schilperoort RA, Nuti MP: Large plasmids of fast-growing Rhizobia: Homology studies and location of structural nitrogen fixation (nif) genes. J Bacteriol 145:1129–1136, 1981.

    Google Scholar 

  36. Rigby WJP, Dieckmann M, Rhodes C, Berg P: Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol 113:237–251, 1977.

    Google Scholar 

  37. Rolfe BG, Gresshoff PM, Shine J: Rapid screening for symbiotic mutants of Rhizobium and white clover. Plant Sci Lett 19:277–284, 1980.

    Google Scholar 

  38. Rosenberg C, Casse-Delbart F, Dusha I, David M, Boucher C: Megaplasmids in the plant-associated bacteria Rhizobium meliloti and Pseudomonas solanacearum. J Bacteriol 150:402–406, 1982.

    Google Scholar 

  39. Ruvkin GB, Ausubel FM: Interspecies homology of nitrogenase genes. Proc Natl Acad Sci USA 77:191–195, 1980.

    Google Scholar 

  40. Scott DB, Ronson CW: Identification and mobilization by cointegrate formation of a nodulation plasmid in Rhizobium trifolii. J Bacteriol 151:36–43, 1982.

    Google Scholar 

  41. Schofield PR, Djordjevic MA, Rolfe BG, Shine J, Watson JM: A molecular linkage map of nitrogenase and nodulation genes in Rhizobium trifolii. Mol Gen Genet 192:459–465, 1983.

    Google Scholar 

  42. Simon R, Priefer U, Puhler A: A broadhost range mobilization system for in vivo genetic engineering: transposon mutagenesis in gram negative bacteria. Biotechnology 1:784–791, 1983.

    Google Scholar 

  43. Stowers MD, Eaglesham ARJ: Physiological and symbiotic characteristics of fast-growing Rhizobium japonicum. Plant Soil 77:3–14, 1984.

    Google Scholar 

  44. Yelton MM, Yang SS, Edie SA, Lim ST: Characterization of an effective salt-tolerant, fast-growing strain of Rhizobium japonicum. J Gen Microbiol 129:1537–1540, 1983.

    Google Scholar 

  45. Zurkowski W, Lorkiewicz: Plasmid-mediated control of nodulation in Rhizobium trifolii. Arch Microbiol 123:195–201, 1979.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Dept. of Genetics, Iowa State University, 50011, Ames, IA, U.S.A.

    Kim Stutzman Engwall & Alan G. Atherly

Authors
  1. Kim Stutzman Engwall
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Alan G. Atherly
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Engwall, K.S., Atherly, A.G. The formation of R-prime deletion mutants and the identification of the symbiotic genes in Rhizobium fredii strain USDA191. Plant Mol Biol 6, 41–51 (1986). https://doi.org/10.1007/BF00021305

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00021305

Keywords

Search

Navigation