Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1985 Nov;82(21):7379–7383. doi: 10.1073/pnas.82.21.7379

Isolation and characterization of nodulation genes from Bradyrhizobium sp. (Vigna) strain IRc 78

John D Noti 1, Brigitta Dudas 1,*, Aladar A Szalay 1,†
PMCID: PMC391348  PMID: 16593624

Abstract

An 11.76-kilobase-pair (kb) segment of DNA from Bradyrhizobium sp. (Vigna) strain IRc 78 that hybridizes to nodulation genes of Rhizobium meliloti strain 41 was isolated. Hybridization of the 11.76-kb DNA fragment to DNA from other Bradyrhizobium species revealed a high degree of sequence conservation in this region. Transfer of the 11.76-kb segment to nodulation-defective (Nod-) mutants of R. meliloti restored their ability to induce nodules on Medicago sativa (alfalfa). Mutants of strain IRc 78 generated by Tn5 mutagenesis of the 11.76-kb segment fell into three classes according to their symbiotic reaction with Vigna unguiculata (cowpea). Class I mutants of strain IRc 78 were unable to induce root-hair curling or to nodulate; class II induced small, ineffective nodules; and class III showed delayed and decreased nodulation with reduction in amount of nitrogen fixed. Furthermore, in contrast to the wild-type strain, class I mutants could not induce nodules on Glycine max (soybean), Cajanus cajan (pigeon pea), or Arachis hypogaea (peanut). This finding suggests a common function of the 11.76-kb region in the infection of host plants by Bradyrhizobium either through root hairs or by “crack entry.”

Keywords: cosmid cloning, Tn5 mutagenesis, functional complementation

Full text

PDF
7379

Images in this article

7380Image
on p.7380
7380Image
on p.7380
7381Image
on p.7381
7381Image
on p.7381
7382Image
on p.7382
7382Image
on p.7382
7382Image
on p.7382

Selected References

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

  1. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Boyer H. W., Roulland-Dussoix D. A complementation analysis of the restriction and modification of DNA in Escherichia coli. J Mol Biol. 1969 May 14;41(3):459–472. doi: 10.1016/0022-2836(69)90288-5. [DOI] [PubMed] [Google Scholar]
  3. 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]
  4. Chinault A. C., Carbon J. Overlap hybridization screening: isolation and characterization of overlapping DNA fragments surrounding the leu2 gene on yeast chromosome III. Gene. 1979 Feb;5(2):111–126. doi: 10.1016/0378-1119(79)90097-0. [DOI] [PubMed] [Google Scholar]
  5. Hadley R. G., Eaglesham A. R., Szalay A. A. Conservation of DNA regions adjacent to nifKDH homologous sequences in diverse slow-growing Rhizobium strains. J Mol Appl Genet. 1983;2(3):225–236. [PubMed] [Google Scholar]
  6. Hanahan D., Meselson M. Plasmid screening at high colony density. Gene. 1980 Jun;10(1):63–67. doi: 10.1016/0378-1119(80)90144-4. [DOI] [PubMed] [Google Scholar]
  7. Maier R. C., Norris H. J. Glassy cell carcinoma of the cervix. Obstet Gynecol. 1982 Aug;60(2):219–224. [PubMed] [Google Scholar]
  8. Meinkoth J., Wahl G. Hybridization of nucleic acids immobilized on solid supports. Anal Biochem. 1984 May 1;138(2):267–284. doi: 10.1016/0003-2697(84)90808-x. [DOI] [PubMed] [Google Scholar]
  9. Morris D. W., Noti J. D., Osborne F. A., Szalay A. A. Plasmid vectors capable of transferring large DNA fragments to yeast. DNA. 1981;1(1):27–36. doi: 10.1089/dna.1.1981.1.27. [DOI] [PubMed] [Google Scholar]
  10. Rigby P. W., 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. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  11. Rosenberg C., Casse-Delbart F., Dusha I., David M., Boucher C. Megaplasmids in the plant-associated bacteria Rhizobium meliloti and Pseudomonas solanacearum. J Bacteriol. 1982 Apr;150(1):402–406. doi: 10.1128/jb.150.1.402-406.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Rossen L., Johnston A. W., Downie J. A. DNA sequence of the Rhizobium leguminosarum nodulation genes nodAB and C required for root hair curling. Nucleic Acids Res. 1984 Dec 21;12(24):9497–9508. doi: 10.1093/nar/12.24.9497. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Ruvkun G. B., Long S. R., Meade H. M., van den Bos R. C., Ausubel F. M. ISRm1: A Rhizobium meliloti insertion sequence that transposes preferentially into nitrogen fixation genes. J Mol Appl Genet. 1982;1(5):405–418. [PubMed] [Google Scholar]
  14. Sternberg N., Tiemeier D., Enquist L. In vitro packaging of a lambda Dam vector containing EcoRI DNA fragments of Escherichia coli and phage P1. Gene. 1977 May;1(3-4):255–280. doi: 10.1016/0378-1119(77)90049-x. [DOI] [PubMed] [Google Scholar]
  15. Török I., Kondorosi E., Stepkowski T., Pósfai J., Kondorosi A. Nucleotide sequence of Rhizobium meliloti nodulation genes. Nucleic Acids Res. 1984 Dec 21;12(24):9509–9524. doi: 10.1093/nar/12.24.9509. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Yang R., Lis J., Wu R. Elution of DNA from agarose gels after electrophoresis. Methods Enzymol. 1979;68:176–182. doi: 10.1016/0076-6879(79)68012-6. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES