Plant and Soil

, Volume 61, Issue 1–2, pp 53–63 | Cite as

Characteristics of nitrogen-fixingKlebsiella oxytoca isolated from wheat roots

  • M. Lutfu Cakmakci
  • H. J. Evans
  • R. J. Seidler


Of 45 fermentative gram negative bacterial isolates examined from wheat roots, three were capable of fixing atmospheric nitrogen as determined by the acetylene reduction technique and by protein contents of cells. A gram negative non-motile facultatively anaerobic bacterial strain capable of N2 fixation was identified asKlebsiella oxytoca ZMK-2.

Optimal growth and N2 fixation occurred at pH 6.5. The optimum temperatures for growth under anaerobic conditions ranged between 30°–37°C. Acetylene reduction by intact cells was strikingly inhibited by 0.1 atm. or greater partial pressure of O2. Furthermore, the accumulation of H2 in the gas phase over cultures ofKlebsiella oxytoca ZMK-2 at partial pressures greater than 0.02 atm. resulted in a striking inhibition in the rate of C2H2 reduction. The addition of suspensions of eitherKlebsiella oxytoca ZMK-2 orAzotobacter vinelandii or a mixed culture of these two organisms to axenic cultures of wheat plants produced no significant increase in plant growth as measured by plant dry weight or nitrogen content of plants.

Key Words

Acetylene reduction Inoculation Klebsiella oxytoca Nitrogen fixation Wheat 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Aho, E. P., Seidler, R. J., Evans, H. J. and Raju, P. N. 1974 Distribution, enumeration and identification of nitrogen fixing bacteria associated with decay in living white fir trees. Phytopathology64, 1413–1420.Google Scholar
  2. 2.
    Alexander, M. 1977 Introduction to Soil Microbiology. John Wiley and Sons, New York.Google Scholar
  3. 3.
    Barber, L. E. and Evans, H. J. 1976 Characterization of a nitrogen fixing bacterial strain from the roots ofDigitaria sanguinalis. Can. J. Microbiol.22, 245–260.Google Scholar
  4. 4.
    Barber, L. E., Russell, S. A. and Evans, H. J. 1979 Inoculation of millet with Azospirillum. Plant and Soil59, 49–57.Google Scholar
  5. 5.
    Bergey's Manual of Determinative Bacteriology 1974 Eds. R. R. Buchanan and N. N. Gibbons. 8th edition. Williams and Wilkins.Google Scholar
  6. 6.
    Conn, H. J. (ed.) 1957 Manual of Microbiological Methods. McGraw-Hill Book Co., New York.Google Scholar
  7. 7.
    Cowan, S. T. 1974 Manual for Identification of Medical Bacteria. 2nd edition. Cambridge University Press.Google Scholar
  8. 8.
    Dobereiner, J., Day, J. M. and Dart, P. J. 1972 Nitrogenase activity in the rhizosphere of sugar cane and some other tropical grasses. Plant and Soil37, 191–196.Google Scholar
  9. 9.
    Dommergues, Y., Balandreau, J., Rinaudo, G. and Weinhard, P. 1973 Non-symbiotic nitrogen fixation in the rhizosphere of rice, maize and different tropical grasses. Soil Biol. Biochem.5, 83–89.Google Scholar
  10. 10.
    Evans, H. J., Campbell, N. E. R. and Hill, S. 1972 Asymbiotic nitrogen fixing bacteria from the surface of nodules and roots of legumes. Can. J. Microbiol.18, 13–21.Google Scholar
  11. 11.
    Goa, J. 1953 A microbiuret method for protein determination of total protein in celebrospinal fluid. Scan. Clin. Lab. Invest.5, 218–222.Google Scholar
  12. 12.
    Klucas, R. 1972 Nitrogen fixation by Klebsiella grown in the presence of oxygen. Can. J. Microb.18, 1845–1850.Google Scholar
  13. 13.
    Line, M. A. and Loutit, M. W. 1971 Non-symbiotic nitrogen fixing organisms from some New Zealand tussock grassland soil. J. Gen. Microbiol.66, 309–318.Google Scholar
  14. 14.
    Mandel, M., Ingambi, L., Mergendahl, J., Dodson, M. L., Jr. and Scheltgen. 1970 Correlation of melting temperature and cesium chloride buoyant density of bacterial deoxyribonucleic acid. J. Bacteriol.101, 333–338.Google Scholar
  15. 15.
    Neilson, A. H. and Sparell. 1976 Acetylene reduction nitrogen fixation by Enterobacteriaceae isolated from paper mill process waters. Appl. Environ. Microbiol.32, 197–205.Google Scholar
  16. 16.
    Raju, P. N., Evans, H. J. and Seidler, R. J. 1972 An asymbiotic nitrogen fixing bacterium from the root environment of corn. Proc. Nat. Acad. Sci.67, 3474–3478.Google Scholar
  17. 17.
    Seidler, R. J., Starr, M. P. and Mandel, M. 1969 Deoxyribonucleic acid (DNA) characterization of Bdellovibrios. J. Bacteriol.100, 787–790.Google Scholar
  18. 18.
    Seidler, R. J., Morrow, J. E. and Bagley, S. T. 1977 Klebsiella in drinking water emanating from red wood tanks. Appl. Environ. Microbiol.33, 893–900.Google Scholar
  19. 19.
    Stenzel, W., Burger, H. and Mannheim, W. 1972 On the systematics and differential diagnosis of the Klebsiella group under special consideration of the so-called oxytocum types. Zentralbl. Bakteriol. Parasitenkd. Infektionskr. Hyg. Abt. 1 Orig. Reihe A219, 193–203.Google Scholar
  20. 20.
    Stickland, L. H. 1951 Determination of small quantities of bacteria by means of the biuret reaction. J. Gen. Microbiol.5, 698–703.Google Scholar
  21. 21.
    Umbreit, W. W., Burris, R. H. and Staufer, J. F. 1957 Manometric Techniques. Burgess Publishing Co., Minneapolis, Minn.Google Scholar
  22. 22.
    Vancura, V. 1964 Root exudates of plants. Plant and Soil21, 231–248.Google Scholar
  23. 23.
    Werner, D., Evans, H. J. and Seidler, R. J. 1974 Facultatively anaerobic nitrogen fixing bacteria from the marine environment. Can. J. Microbiol.20, 59–64.Google Scholar
  24. 24.
    Wilcockson, J. and Werner, D. 1976 Nitrogenase activity by Klebsiella and Rhizobium on solid substrate exposed to air. Ber. Dtsch. Bot. Ges.89, 587.Google Scholar

Copyright information

© Martinus Nijhoff/Dr W. Junk Publishers 1981

Authors and Affiliations

  • M. Lutfu Cakmakci
    • 1
  • H. J. Evans
    • 2
  • R. J. Seidler
    • 3
  1. 1.Department of Agricultural Microbiology, Faculty of AgricultureUniversity of AnkaraTurkey
  2. 2.Laboratory for Nitrogen Fixation ResearchOregon State UniversityCorvallis
  3. 3.Department of MicrobiologyOregon State UniversityCorvallis

Personalised recommendations