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Archives of Microbiology

, Volume 142, Issue 3, pp 229–233 | Cite as

Assimilation of exogenous and dinitrogen-derived13NH 4 + byAnabaena azollae separated fromAzolla caroliniana Willd

  • John C. Meeks
  • Nisan Steinberg
  • Cecillia M. Joseph
  • Carol S. Enderlin
  • Paul A. Jorgensen
  • Gerald A. Peters
Original Papers

Abstract

Anabaena azollae was isolated fromAzolla caroliniana by the “gentle roller” method and differential centrifugation. Incubation of suchAnabaena preparations for 10 min with [13N]N2 resulted in the formation of four radioactive compounds; ammonium, glutamine, glutamate and alanine. Ammonium accounted for 66% of the total radioactivity recovered and 58% of the ammonium was in an extracellular fraction. Since essentially no extracellular13N-labeled organic compounds were found, it appears that ammonium is the compound most probably made available toAzolla during dinitrogen-dependent growth of the association.

The kinetics of incorporation of exogenous13NH 4 + into glutamine and glutamate were characteristic of a precursor (glutamine)-product (glutamate) relationship and consistent with assimilation by the glutamine synthetase-glutamate synthase pathway. The results of experiments using the glutamine synthetase inhibitor, methionine sulfoximine, the glutamate synthase inhibitor, diazo-oxonorleucine, and increasing the ammonium concentration to greater than 1 mM, provided evidence for assimilation primarily by the glutamine synthetase-glutamate synthase pathway with little or no contribution from biosynthetic glutamate dehydrogenase.

While showing that N2 fixation and NH 4 + assimilation were not tightly coupled metabolic processes in symbioticAnabaena, these results reflect a composite picture and do not indicate the extent to which ammonium assimilatory enzymes might be regulated in filaments associated with specific stages in theAzolla-Anabaena developmental profile.

Key words

Ammonium assimilation Excretion Anabaena azollae Azolla caroliniana Cyanobacteria Glutamine Glutamate formation Nitrogen fixation Symbiosis 

Non-standard abbreviations

DON

6-Diazo-5-oxo-l-norleucine

GDH

glutamate dehydrogenase

GOGAT

glutamate synthase

GS

glutamine synthetase

MSX

l-methionine-Dl-sulfoximine

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Copyright information

© Springer-Verlag 1985

Authors and Affiliations

  • John C. Meeks
    • 1
  • Nisan Steinberg
    • 1
  • Cecillia M. Joseph
    • 1
  • Carol S. Enderlin
    • 1
  • Paul A. Jorgensen
    • 1
  • Gerald A. Peters
    • 2
  1. 1.Department of BacteriologyUniversity of CaliforniaDavisUSA
  2. 2.Battelle-C.F. Kettering Research LaboratoryYellow SpringsUSA

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