Skip to content
1887

Microbiology Society logo Microbiology

Volume 133, Issue 1

Conditions for Mutagenesis of the Nitrogen-fixing Cyanobacterium Free

Abstract

Chemically induced mutation in the cyanobacterium was studied using resistance to the pyrimidine analogue 5′-fluorocytosine as a genetic marker which can be selected positively. Cytosine is metabolized through uracil and the UMP pyrophosphorylase ‘salvage’ pathway in this photoautotroph, as it is in enteric bacteria. Treatment with various concentrations of -methyl-′-nitro--nitrosoguanidine (MNNG) gave the highest frequencies of 5FC-resistant mutants when lethality approximated 99%, irrespective of the exposure time or mutagen concentration. The pH of the incubation medium strongly influenced mutation; exposure to MNNG at pH 6·0 yielded 13-fold higher frequencies of mutants than at pH 7·5. The greatest frequency of resistant cells was found after cultures had undergone six or more doublings following mutagenesis. The mutation frequencies obtained by treatment with MNNG were approximately 4- and 25-fold higher than those after exposure under empirically defined conditions to diethyl sulphate or nitrous acid, respectively, and 1·4 × 10-fold higher than the frequency of spontaneous mutation. Neither chloramphenicol-inhibited nor caffeine-sensitive systems capable of repairing MNNG-induced DNA damage were observed.

  • Received:
  • Published Online:
© Society for General Microbiology 1987
Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-133-1-111
1987-01-01
2025-12-11

Metrics

Loading full text...

Full text loading...

/deliver/fulltext/micro/133/1/mic-133-1-111.html?itemId=/content/journal/micro/10.1099/00221287-133-1-111&mimeType=html&fmt=ahah

References

  1. Adelberg E. A., Mandel M., Chen G. C. 1965; Optimal conditions for mutagenesis by N-methyl-N´- nitro-N-nitrosoguanidine in Escherichia coli. Biochemical and Biophysical Research Communications 18:788–795
     [Google Scholar]
  2. Allen M. B., Arnon D. I. 1955; Studies on nitrogen-fixing blue-green algae. I. Growth and nitrogen fixation by Anabaena cylindrica Lemm. Plant Physiology 30:366–372
     [Google Scholar]
  3. Astier C., Joset-Espardellier F., Meyer I. 1979; Conditions for mutagenesis in the cyanobacterium Aphanocapsa 6714. Influence of repair phenomena. Archives of Microbiology 120:93–96
     [Google Scholar]
  4. Bautz E., Freese E. 1960; In vivo action of alkylating mutagens. Proceedings of the National Academy of Sciences of the United States of America 46:1585–1594
     [Google Scholar]
  5. Beck C. F., Ingraham J. I., Neuhard J., Thomassen E. 1972; Metabolism of pyrimidines and pyrimidine nucleosides by Salmonella typhimurium. Journal of Bacteriology 110:219–228
     [Google Scholar]
  6. Braun A. C., Wood H. N. 1962; On the activation of certain essential biosynthetic systems in cells of Vinca rosea. Proceedings of the National Academy of Sciences of the United States of America 48:1776–1782
     [Google Scholar]
  7. Buzby J. S., Porter R. D., Stevens S. E. Jr 1983; Plasmid transformation in Agmenellum quad- ruplicatum PR-6: construction of biphasic plasmids and characterization of their transformation properties. Journal of Bacteriology 154:1446–1450
     [Google Scholar]
  8. Chapman J. S. 1984 Parameters influencing the isolation of auxotrophs of Anabaena variabilis PhD thesis University of California, Davis, USA:
     [Google Scholar]
  9. Chapman J. S., Meeks J. C. 1983; Glutamine and glutamate transport by Anabaena variabilis. Journal of Bacteriology 156:122–129
     [Google Scholar]
  10. Currier T. C., Wolk C. P. 1978; Regulation of uridylic acid biosynthesis in the cyanobacterium Anabaena variabilis. Journal of Bacteriology 136:682–687
     [Google Scholar]
  11. Currier T. C., Haury J. F., Wolk C. P. 1977; Isolation and characterization of auxotrophs of a filamentous cyanobacterium. Journal of Bacteriology 129:1556–1562
     [Google Scholar]
  12. Delaney S. F., Carr N. G. 1975; Temporal genetic mapping in the blue-green alga Anacystis nidulans using ethylmethane sulphonate. Journal of General Microbiology 88:259–268
     [Google Scholar]
  13. Doolittle W. F. 1979; The cyanobacterial genome, its expression and the control of that expression. Advances in Microbial Physiology 20:1–102
     [Google Scholar]
  14. Guerola N., Ingraham J. L., Cerda-Olmedo E. 1971; Induction of closely linked multiple mutation by N-methyl-N′-nitro-N-nitrosoguanidine. Nature New Biology 230:122–125
     [Google Scholar]
  15. Herdman M. 1982; Evolution and genetic properties of cyanobacterial genomes. In The Biology of Cyanobacteria pp. 263–305 Carr N. G., Whitton B. A. Edited by Oxford: Blackwell Scientific Publications;
     [Google Scholar]
  16. Herdman M., Carr N. G. 1972; The isolation and characterization of mutant strains of the blue-green alga Anacystis nidulans. Journal of General Microbiology 70:213–220
     [Google Scholar]
  17. Herdman M., Delaney S. F., Carr N. G. 1980; Mutation of the cyanobacterium Anacystis nidulans (Synechococcus PCC 6301): improved conditions for the isolation of auxotrophs. Archives of Microbiology 124:177–184
     [Google Scholar]
  18. Kato T. 1977; Effects of chloramphenicol and caffeine on postreplication repair in uvrA−umuC− and uvrA− recF− strains of Escherichia coli K12. Molecular and General Genetics 156:115–120
     [Google Scholar]
  19. Kimball R. F., Setlow J. K. 1974; Mutation fixation in MNNG-treated Haemophilus influenzae as determined by transformation. Mutation Research 22:1–14
     [Google Scholar]
  20. Neuhard J., Ingraham J. L. 1968; Mutants of Salmonella typhimurium requiring cytosine or cytidine for growth. Journal of Bacteriology 95:2431–2434
     [Google Scholar]
  21. O’Donovan G. A., Neuhard J. 1970; Pyrimidine metabolism in microorganisms. Bacteriological Reviews 34:278–343
     [Google Scholar]
  22. Rippka R., Deruelles J., Waterbury J., Herdman M., Stanier R. 1979; Generic assignments, strain histories and properties of pure cultures of cyanobacteria. Journal of General Microbiology 111:1–61
     [Google Scholar]
  23. Shaffer P. W., Lockau W., Wolk C. P. 1978; Isolation of mutants of the cyanobacterium, Anabaena variabilis, impaired in photoautotrophy. Archives of Microbiology 117:215–219
     [Google Scholar]
  24. Singh R. N., Kashyap K. N. 1977; Induction of mutations in the blue-green alga Plectonema boryanum. Mutation Research 43:37–44
     [Google Scholar]
  25. Sklar R. 1978; Enhancement of nitrosoguanidine mutagenesis by chloramphenicol in Escherichia coli K12. Journal of Bacteriology 136:460–462
     [Google Scholar]
  26. Stacey G., Van Baalen C., Tabita F. R. 1979; Nitrogen and ammonia assimilation in the cyanobacteria: regulation of glutamine synthetase. Archives of Biochemistry and Biophysics 194:457–467
     [Google Scholar]
  27. Stevens S. E. Jr Van Baalen C. 1969; N-methyl-N′-nitro-N-nitrosoguanidine as a mutagen for blue-green algae: evidence for repair. Journal of Phycology 5:136–139
     [Google Scholar]
  28. Tang T., Asato Y. 1978; Ultraviolet light induction and photoreactivation of thymine dimers in Anacystis nidulans. Archives of Microbiology 119:193–197
     [Google Scholar]
  29. Wilcox M., Mitchison G. J., Smith R. J. 1975; Spatial control of differentiation in the blue-green alga Anabaena cylindrica. In Microbiology - 1975 pp. 453–463 Schlessinger D. Edited by Washington, DC: American Society for Microbiology;
     [Google Scholar]
  30. Wolk C. P., Wojciuch E. 1973; Simple methods for plating single vegetative cells of, and for replica plating, filamentous blue-green algae. Archiv für Mikrobiologie 91:91–95
     [Google Scholar]
  31. Wolk C. P., Vonshak A., Kehoe P., Elhai J. 1984; Construction of shuttle vectors capable of conjugative transfer from Escherichia coli to nitrogen-fixing filamentous cyanobacteria. Proceedings of the National Academy of Sciences of the United States of America 81:1516–1565
     [Google Scholar]
  32. Zimmerman F. K. 1977; Genetic effects of nitrous acid. Mutation Research 39:127–148
     [Google Scholar]
/content/journal/micro/10.1099/00221287-133-1-111
Loading
Conditions for Mutagenesis of the Nitrogen-fixing Cyanobacterium Anabaena variabilis
Microbiology 133, 111 (1987); https://doi.org/10.1099/00221287-133-1-111
/content/journal/micro/10.1099/00221287-133-1-111
/content/journal/micro/10.1099/00221287-133-1-111
Loading

Data & Media loading...

Most cited Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An error occurred
Approval was partially successful, following selected items could not be processed due to error