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. 1973 Apr;114(1):164–168. doi: 10.1128/jb.114.1.164-168.1973

Selection of Respiratory Mutants of Neurospora crassa

David L Edwards 1, Fabia Kwiecinski 1, Jonathan Horstmann 1
PMCID: PMC251752  PMID: 4349028

Abstract

A method is described that permits the isolation of mutants that are defective in mitochondrial respiration. The techniques of inositol-less death and overlay with 2,3,5-triphenyl-2H-tetrazolium chloride are utilized to select for mutant colonies. Colonies that survive inositol-less death and fail to reduce the tetrazolium dye are then tested polarographically for cyanide-sensitive respiration. A preliminary characterization of three mutants obtained by this method is presented. The mutants have been characterized by their cyanide-sensitive respiration rate, growth rate, the state III respiration rate of isolated mitochondria, inhibition of the respiration of isolated mitochondria by cyanide and antimycin A, and cytochrome spectra. All of the mutants described differ from the parent strain in some of these aspects.

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Selected References

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

  1. Bertrand H., Pittenger T. H. Cytoplasmic Mutants Selected from Continuously Growing Cultures of NEUROSPORA CRASSA. Genetics. 1969 Mar;61(3):643–659. doi: 10.1093/genetics/61.3.643. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bertrand H., Pittenger T. H. Isolation and classification of extranuclear mutants of Neurospora crassa. Genetics. 1972 Aug;71(4):521–533. doi: 10.1093/genetics/71.4.521. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Eakin R. T., Mitchell H. K. Alterations of the respiratory system of Neurospora crassa by the mi-1 mutation. J Bacteriol. 1970 Oct;104(1):74–78. doi: 10.1128/jb.104.1.74-78.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Edwards D. L., Woodward D. O. An altered cytochrome oxidase in a cytoplasmic mutant of Neurospora. FEBS Lett. 1969 Aug;4(3):193–196. doi: 10.1016/0014-5793(69)80232-2. [DOI] [PubMed] [Google Scholar]
  5. Flavell R. B., Fincham J. R. Acetate-nonutilizing mutants of Neurospora rassa. II. Biochemical deficiencies and the roles of certain enzymes. J Bacteriol. 1968 Mar;95(3):1063–1068. doi: 10.1128/jb.95.3.1063-1068.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Flavell R. B., Fincham J. R. Acetate-onutilizing mutants of Neurospora crassa. I. Mutant isolation, complementation studies, and linkage relationships. J Bacteriol. 1968 Mar;95(3):1056–1062. doi: 10.1128/jb.95.3.1056-1062.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Flavell R. B., Woodward D. O., Edwards D. L. Nucleocytoplasmic interactions in the control of mitochondrial structure and function in Neurospora. Symp Soc Exp Biol. 1970;24:55–69. [PubMed] [Google Scholar]
  8. Garnjobst L., Wilson J. F., Tatum E. L. Studies on a cytoplasmic character in Neurospora crassa. J Cell Biol. 1965 Aug;26(2):413–425. doi: 10.1083/jcb.26.2.413. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gillie O. J. Methods for the study of nuclear and cytoplasmic variation in respiratory activity of Neurospora crassa, and the discovery of three new genes. J Gen Microbiol. 1970 Jun;61(3):379–395. doi: 10.1099/00221287-61-3-379. [DOI] [PubMed] [Google Scholar]
  10. HASKINS F. A., TISSIERES A., MITCHELL H. K., MITCHELL M. B. Cytochromes and the succinic acid oxidase system of poky strains of Neurospora. J Biol Chem. 1953 Feb;200(2):819–826. [PubMed] [Google Scholar]
  11. 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]
  12. Lambowitz A. M., Slayman C. W. Cyanide-resistant respiration in Neurospora crassa. J Bacteriol. 1971 Dec;108(3):1087–1096. doi: 10.1128/jb.108.3.1087-1096.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Lambowitz A. M., Slayman C. W., Slayman C. L., Bonner W. D., Jr The electron transport components of wild type and poky strains of Neurospora crassa. J Biol Chem. 1972 Mar 10;247(5):1536–1545. [PubMed] [Google Scholar]
  14. McDougall K. J., Pittenger T. H. A cytoplasmic variant of Neurospora crassa. Genetics. 1966 Aug;54(2):551–565. doi: 10.1093/genetics/54.2.551. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Mitchell M. B., Mitchell H. K. A Case of "Maternal" Inheritance in Neurospora Crassa. Proc Natl Acad Sci U S A. 1952 May;38(5):442–449. doi: 10.1073/pnas.38.5.442. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Schonbaum G. R., Bonner W. D., Jr, Storey B. T., Bahr J. T. Specific inhibition of the cyanide-insensitive respiratory pathway in plant mitochondria by hydroxamic acids. Plant Physiol. 1971 Jan;47(1):124–128. doi: 10.1104/pp.47.1.124. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. TISSIERES A., MITCHELL H. K. Cytochromes and respiratory activities in some slow growing strains of Neurospora. J Biol Chem. 1954 May;208(1):241–249. [PubMed] [Google Scholar]
  18. TISSIERES A., MITCHELL H. K., HASKINS F. A. Studies on the respiratory system of the poky strain of Neurospora. J Biol Chem. 1953 Nov;205(1):423–433. [PubMed] [Google Scholar]
  19. Weiss H., von Jagow G., Klingenberg M., Bücher T. Characterization of Neurospora crassa mitochondria prepared with a grind-mill. Eur J Biochem. 1970 May 1;14(1):75–82. doi: 10.1111/j.1432-1033.1970.tb00263.x. [DOI] [PubMed] [Google Scholar]

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