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. 1977 Nov;60(5):771–774. doi: 10.1104/pp.60.5.771

Isolation of Bacteria, Transforming Bacteria, and Bacteroids from Soybean Nodules 1

Te May Ching a, Saundra Hedtke a, William Newcomb b
PMCID: PMC542712  PMID: 16660182

Abstract

Postnuclei supernatant of soybean (Glycine max cv. Chippewa 64) nodule homogenate was fractionated by stepwise sucrose density gradient centrifugation into supernatant, endoplasmic reticulum and mitochondria, and three distinct bands with 1.22, 1.25, and 1.27 g/cm3 of peak density. Based on their enzymic activities, composition of electron transport components, and ultrastructural characteristics, the lightest band appears to be the mature bacteroids; the intermediate band the transforming bacteria; and the heaviest, the bacteria. The isolation procedure separates nodule symbionts into different functional and developmental fractions, and it may be a valuable tool for studies involving development, regulation, and senescence of bacteroids in the nodule.

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Selected References

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

  1. Appleby C. A. Electron transport systems of Rhizobium japonicum. I. Haemoprotein P-450, other CO-reactive pigments, cytochromes and oxidases in bacteroids from N2-fixing root nodules. Biochim Biophys Acta. 1969 Jan 14;172(1):71–87. doi: 10.1016/0005-2728(69)90093-0. [DOI] [PubMed] [Google Scholar]
  2. Appleby C. A. Electron transport systems of Rhizobium japonicum. II. Rhizobium haemoglobin, cytochromes and oxidases in free-living (cultured) cells. Biochim Biophys Acta. 1969 Jan 14;172(1):88–105. doi: 10.1016/0005-2728(69)90094-2. [DOI] [PubMed] [Google Scholar]
  3. Appleby C. A., Turner G. L., Macnicol P. K. Involvement of oxyleghaemoglobin and cytochrome P-450 in an efficient oxidative phosphorylation pathway which supports nitrogen fixation in Rhizobium. Biochim Biophys Acta. 1975 Jun 17;387(3):461–474. doi: 10.1016/0005-2728(75)90086-9. [DOI] [PubMed] [Google Scholar]
  4. Bergersen F. J., Turner G. L., Appleby C. A. Studies of the physiological role of leghaemoglobin in soybean root nodules. Biochim Biophys Acta. 1973 Jan 18;292(1):271–282. doi: 10.1016/0005-2728(73)90271-5. [DOI] [PubMed] [Google Scholar]
  5. Doern G. V., Buckmire F. L. Ultrastructural characterization of capsulated Haemophilus influenzae type b and two spontaneous nontypable mutants. J Bacteriol. 1976 Jul;127(1):523–535. doi: 10.1128/jb.127.1.523-535.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Evans H. J., Koch B., Klucas R. Preparation of nitrogenase from nodules and separation into components. Methods Enzymol. 1972;24:470–476. doi: 10.1016/0076-6879(72)24092-7. [DOI] [PubMed] [Google Scholar]
  7. Gerhardt B., Beevers H. Influence of sucrose on protein determination by the Lowry procedure. Anal Biochem. 1968 Aug;24(2):337–339. doi: 10.1016/0003-2697(68)90187-5. [DOI] [PubMed] [Google Scholar]
  8. Goodchild D. J., Bergersen F. J. Electron microscopy of the infection and subsequent development of soybean nodule cells. J Bacteriol. 1966 Jul;92(1):204–213. doi: 10.1128/jb.92.1.204-213.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Huang A. H. Comparative studies of glyoxysomes from various Fatty seedlings. Plant Physiol. 1975 May;55(5):870–874. doi: 10.1104/pp.55.5.870. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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]
  11. Tjepkema J., Evans H. J. Nitrogen fixation by free-living Rhizobium in a defined liquid medium. Biochem Biophys Res Commun. 1975 Jul 22;65(2):625–628. doi: 10.1016/s0006-291x(75)80192-6. [DOI] [PubMed] [Google Scholar]
  12. VENABLE J. H., COGGESHALL R. A SIMPLIFIED LEAD CITRATE STAIN FOR USE IN ELECTRON MICROSCOPY. J Cell Biol. 1965 May;25:407–408. doi: 10.1083/jcb.25.2.407. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Wittenberg J. B. Facilitated oxygen diffusion. The role of leghemoglobin in nitrogen fixation by bacteroids isolated from soybean root nodules. J Biol Chem. 1974 Jul 10;249(13):4057–4066. [PubMed] [Google Scholar]
  14. Wong P. P., Evans H. J. Poly-beta-hydroxybutyrate Utilization by Soybean (Glycine max Merr.) Nodules and Assessment of Its Role in Maintenance of Nitrogenase Activity. Plant Physiol. 1971 Jun;47(6):750–755. doi: 10.1104/pp.47.6.750. [DOI] [PMC free article] [PubMed] [Google Scholar]

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