Archives of Microbiology

, Volume 151, Issue 6, pp 469–474 | Cite as

Evidence that the barrier to the penetration of oxygen into heterocysts depends upon two layers of the cell envelope

  • Marcia A. Murry
  • C. Peter Wolk
Original Papers


Mutants of Anabaena sp. PCC 7120 with O2-sensitive acetylene-reducing activity were studied to identify envelope components that contribute to the barrier limiting diffusion of oxygen into the heterocyst. Mutant strain EF114, deficient in a heterocyst-specific glycolipid, reduced acetylene only under strictly anaerobic conditions. Analysis of in vivo O2 uptake as a function of dissolved pO2 showed that EF114 has lost the low affinity, diffusion-limited respiratory component associated with heterocysts in wild-type filaments. The low affinity respiratory activity was also lost in EF116, a mutant in which the cohesiveness of the outer polysaccharide layer was reduced. Restoration of aerobic nitrogen fixation in a spontaneous revertant of EF116 and in a strain complemented with cosmid 41E11 was associated with restoration of the low affinity component of respiratory activity. The results provide evidence that the barrier to diffusion of gas into heterocysts depends upon both the glycolipid layer and the polysaccharide layer of the heterocyst envelope.

Key words

Anabaena Heterocysts Nitrogenase Oxygen-protection Mutants 





Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Allen MB, Arnon DI (1955) Studies on nitrogen-fixing blue-green algae. I. Growth and nitrogen fixation by Anabaena cylindrica Lemm. Plant Physiol 30:366–372Google Scholar
  2. Allen MB, Smith AJ (1969) Nitrogen chlorosis in blue-green algae. Arch Mikrobiol 69:114–120Google Scholar
  3. Almon H, Böhme H (1980) Components and activity of the photosynthetic electron transport system of intact heterocysts isolated from the blue-green alga Nostoc muscorum. Biochim Biophys Acta 592:113–120Google Scholar
  4. Apte SK, Rowell P, Stewart WDP (1978) Electron donation to ferredoxin in heterocysts of the N2-fixing alga Anabaena cylindrica. Proc Roy Soc Lond B 200:1–25Google Scholar
  5. Bottomley PJ, Stewart WDP (1977) ATP and nitrogenase activity in nitrogen-fixing heterocystous blue-green algae. New Phytol 79:625–638Google Scholar
  6. Donze M, Haveman J, Schiereck P (1971) Absence of photosystem II in heterocysts of the blue-green alga Anabaena. Biochim Biophys Acta 256:157–161Google Scholar
  7. Duggan JX, Anderson LE (1975) Light regulation of enzyme activity in Anacystis nidulans (Richt.) Planta 122:293–297Google Scholar
  8. Ernst A, Kirschenlohr H, Diez J, Böger P (1984) Glycogen content and nitrogenase activity in Anabaena variabilis. Arch Microbiol 140:120–125Google Scholar
  9. Fay P (1976) Factors influencing dark nitrogen fixation in a bluegreen alga. Appl Environ Microbiol 31:376–379Google Scholar
  10. Hallenbeck PC, Kostel PJ, Benemann JR (1979) Purification and properties of nitrogenase from the cyanobacterium Anabaena cylindrica. Eur J Biochem 98:275–284Google Scholar
  11. Haury JF, Wolk CP (1978) Classes of Anabaena variabilis mutants with oxygen-sensitive nitrogenase activity. J Bacteriol 136: 688–692Google Scholar
  12. Jensen BB, Cox RP (1983) Effect of oxygen concentration on dark nitrogen fixation and respiration in cyanobacteria. Arch Microbiol 135:287–292Google Scholar
  13. Lambein F, Wolk CP (1973) Structural studies on the glycolipids from the envelope of the heterocyst of Anabaena cylindrica. Biochem 12:791–798Google Scholar
  14. Lang NJ, Fay P (1971) The heterocysts of blue-green algae. II. Details of ultrastructure. Proc Roy Soc Lond B 178:193–203Google Scholar
  15. Lommen PW, Schwintzer CR, Yocum CS, Gates DM (1971) A model describing photosynthesis in terms of gas diffusion and enzyme kinetics. Planta 98:195–220Google Scholar
  16. Mackinney G (1941) Absorption of light by chlorophyll solutions. J Biol Chem 140:315–322Google Scholar
  17. Murry MA, Olafsen AO, Benemann JR (1981) Oxidation of diaminobenzidine in the heterocysts of Anabaena cylindrica. Curr Microbiol 6:201–206Google Scholar
  18. Murry MA, Horne AJ, Benemann JR (1984) Physiological studies of oxygen protection mechanisms in the heterocysts of Anabaena cylindrica. Appl Environ Microbiol 47:449–454Google Scholar
  19. Pelroy RA, Rippka R, Stanier R (1972) Metabolism of glucose by unicellular blue-green algae. Arch Mikrobiol 87:303–322Google Scholar
  20. Peterson RB, Burris RH (1976) Properties of heterocysts isolated with colloidal silica. Arch Microbiol 108:35–40Google Scholar
  21. Peterson RB, Burris RH (1978) Hydrogen metabolism in isolated heterocysts of Anabaena 7120. Arch Microbiol 116:125–132Google Scholar
  22. Peterson RB, Wolk CP (1978) Localization of an uptake hydrogense in Anabaena. Plant Physiol 61:688–691Google Scholar
  23. Pienkos PT, Bodmer S, Tabita FR (1983) Oxygen inactivation and recovery of nitrogenase activity in cyanobacteria. J Bacteriol 153:182–190Google Scholar
  24. Rhodes KS (1981) Oxygen sensitivity of nitrogen fixation in the cyanobacterium Anabaenopsis arnoldii. Ph. D. Thesis, University of Michigan, Ann ArborGoogle Scholar
  25. Rippka R, Deruelles J, Waterbury JB, Herdman M, Stanier RY (1979) Generic assignments, strain histories and properties of pure cultures of cyanobacteria. J Gen Microbiol 111:1–61Google Scholar
  26. Rippka R, Waterbury JB (1977) The synthesis of nitrogenase by non-heterocystous cyanobacteria. FEMS Microbiol Lett 2: 83–86Google Scholar
  27. Robson RL, Postgate JR (1980) Oxygen and hydrogen in biological nitrogen fixation. Ann Rey Microbiol 34:183–207Google Scholar
  28. Scherer S, Böger P (1982) Respiration of blue-green algae in the light. Arch Microbiol 132:329–332Google Scholar
  29. Tel-Or E, Stewart WDP (1976) Photosynthetic electron transport, ATP synthesis, and nitrogenase activity in isolated heterocysts of Anabaena cylindrica. Biochim Biophys Acta 423:189–195Google Scholar
  30. Tel-Or E, Stewart WDP (1977) Photosynthetic components and activities of nitrogen-fixing isolated heterocysts of Anabaena cylindrica. Proc R Soc Lond B 198:61–86Google Scholar
  31. Walsby A (1985) The permeability of heterocysts to the gases nitrogen and oxygen. Proc R Soc Lond B 226:345–366Google Scholar
  32. Winkenbach F, Wolk CP (1973) Activities of enzymes of the oxidative and reductive pentose phosphate pathways in heterocysts of a blue-green alga. Plant Physiol 52:480–483Google Scholar
  33. Wolk CP (1973) Physiology and cytological chemistry of blue-green algae. Bacteriol Rev 37:32–101Google Scholar
  34. Wolk CP, Cai Y, Cardemil L, Flores E, Hohn B, Murry M, Schmetterer G, Schrautemeier B, Wilson R (1988) Isolation and complementation of mutants of Anabaena sp. PCC 7120 that are unable to grow aerobically on dinitrogen. J Bacteriol 170:1239–1244Google Scholar

Copyright information

© Springer-Verlag 1989

Authors and Affiliations

  • Marcia A. Murry
    • 1
    • 2
  • C. Peter Wolk
    • 1
    • 2
  1. 1.MSU-DOE Plant Research LaboratoryMichigan State UniversityE. LansingUSA
  2. 2.Department of Botany and Plant PathologyMichigan State UniversityE. LansingUSA

Personalised recommendations