Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1986 Apr;166(1):301–305. doi: 10.1128/jb.166.1.301-305.1986

Isolation and nitrogen-fixing activity of Frankia sp. strain CpI1 vesicles.

N A Noridge, D R Benson
PMCID: PMC214592  PMID: 2870056

Abstract

Under N2-fixing conditions in aerobic culture and in symbiosis, frankiae produce spherical, multicellular structures that have been called vesicles. The vesicles have been proposed as the site of nitrogen fixation. We isolated vesicles by using density centrifugation in a single-step sucrose gradient. Vesicles migrated out of 50% (wt/vol) sucrose and banded at the 40 to 50% sucrose interface; they were intact, as assessed by transmission electron microscopy, and were free of hyphal contamination. Specific activities of nitrogenase in vesicles prepared anaerobically were up to 100-fold greater than the specific activity of the largely hyphal pellet, depending on the recovery of vesicles. All of the activity in the pellet could be accounted for by the number of vesicles present in the pellet. Glutamine synthetase activity in crude extracts of vesicles was extremely low.

Full text

PDF
301

Images in this article

303Image
on p.303
304Image
on p.304

Selected References

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

  1. Baker D., Newcomb W., Torrey J. G. Characterization of an ineffective actinorhizal microsymbiont, Frankia sp. EuI1 (Actinomycetales). Can J Microbiol. 1980 Sep;26(9):1072–1089. doi: 10.1139/m80-180. [DOI] [PubMed] [Google Scholar]
  2. Bender R. A., Janssen K. A., Resnick A. D., Blumenberg M., Foor F., Magasanik B. Biochemical parameters of glutamine synthetase from Klebsiella aerogenes. J Bacteriol. 1977 Feb;129(2):1001–1009. doi: 10.1128/jb.129.2.1001-1009.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Callaham D., Deltredici P., Torrey J. G. Isolation and Cultivation in vitro of the Actinomycete Causing Root Nodulation in Comptonia. Science. 1978 Feb 24;199(4331):899–902. doi: 10.1126/science.199.4331.899. [DOI] [PubMed] [Google Scholar]
  4. Chaykin S. Assay of nicotinamide deamidase. Determination of ammonia by the indophenol reaction. Anal Biochem. 1969 Oct 1;31(1):375–382. doi: 10.1016/0003-2697(69)90278-4. [DOI] [PubMed] [Google Scholar]
  5. Fleming H., Haselkorn R. Differentiation in Nostoc muscorum: nitrogenase is synthesized in heterocysts. Proc Natl Acad Sci U S A. 1973 Oct;70(10):2727–2731. doi: 10.1073/pnas.70.10.2727. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Fontaine M. S., Lancelle S. A., Torrey J. G. Initiation and ontogeny of vesicles in cultured Frankia sp. strain HFPArI3. J Bacteriol. 1984 Dec;160(3):921–927. doi: 10.1128/jb.160.3.921-927.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gauthier D., Diem H. G., Dommergues Y. In vitro nitrogen fixation by two actinomycete strains isolated from casuarina nodules. Appl Environ Microbiol. 1981 Jan;41(1):306–308. doi: 10.1128/aem.41.1.306-308.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  9. Lalonde M., Knowles R. Ultrastructure of the Alnus crispa var. mollis Fern. root nodule endophyte. Can J Microbiol. 1975 Jul;21(7):1058–1080. doi: 10.1139/m75-157. [DOI] [PubMed] [Google Scholar]
  10. Lambein F., Wolk C. P. Structural studies on the glycolipids from the envelope of the heterocyst of Anabaena cylindrica. Biochemistry. 1973 Feb 27;12(5):791–798. doi: 10.1021/bi00729a002. [DOI] [PubMed] [Google Scholar]
  11. Ludwig R. A. Rhizobium free-living nitrogen fixation occurs in specialized nongrowing cells. Proc Natl Acad Sci U S A. 1984 Mar;81(5):1566–1569. doi: 10.1073/pnas.81.5.1566. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Mazzucco C. E., Benson D. R. [14C]methylammonium transport by Frankia sp. strain CpI1. J Bacteriol. 1984 Nov;160(2):636–641. doi: 10.1128/jb.160.2.636-641.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Murry M. A., Fontaine M. S., Tjepkema J. D. Oxygen protection of nitrogenase in Frankia sp. HFPArI3. Arch Microbiol. 1984 Oct;139(2-3):162–166. doi: 10.1007/BF00401993. [DOI] [PubMed] [Google Scholar]
  14. Robson R. L., Postgate J. R. Oxygen and hydrogen in biological nitrogen fixation. Annu Rev Microbiol. 1980;34:183–207. doi: 10.1146/annurev.mi.34.100180.001151. [DOI] [PubMed] [Google Scholar]
  15. Schubert K. R., Coker G. T. Ammonia Assimilation in Alnus glutinosa and Glycine max: SHORT-TERM STUDIES USING [N]AMMONIUM. Plant Physiol. 1981 Apr;67(4):662–665. doi: 10.1104/pp.67.4.662. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Streicher S. L., Tyler B. Regulation of glutamine synthetase activity by adenylylation in the Gram-positive bacterium Streptomyces cattleya. Proc Natl Acad Sci U S A. 1981 Jan;78(1):229–233. doi: 10.1073/pnas.78.1.229. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Thomas J., Meeks J. C., Wolk C. P., Shaffer P. W., Austin S. M. Formation of glutamine from [13n]ammonia, [13n]dinitrogen, and [14C]glutamate by heterocysts isolated from Anabaena cylindrica. J Bacteriol. 1977 Mar;129(3):1545–1555. doi: 10.1128/jb.129.3.1545-1555.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Tjepkema J. D., Ormerod W., Torrey J. G. Factors affecting vesicle formation and acetylene reduction (nitrogenase activity) in Frankia sp. CpI1. Can J Microbiol. 1981 Aug;27(8):815–823. doi: 10.1139/m81-126. [DOI] [PubMed] [Google Scholar]
  19. Wolk C. P., Thomas J., Shaffer P. W., Austin S. M., Galonsky A. Pathway of nitrogen metabolism after fixation of 13N-labeled nitrogen gas by the cyanobacterium, Anabaena cylindrica. J Biol Chem. 1976 Aug 25;251(16):5027–5034. [PubMed] [Google Scholar]

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

RESOURCES