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. 1970 Jan;101(1):1–8. doi: 10.1128/jb.101.1.1-8.1970

Replacement Sporulation of Bacillus subtilis 168 in a Chemically Defined Medium

Robert F Ramaley 1, Linda Burden 1
PMCID: PMC250444  PMID: 4983649

Abstract

A replacement sporulation technique (i.e., the sporulation of vegetative cells upon suspension in an appropriate medium) has been developed for Bacillus subtilis 168 (a transformable Marburg strain of B. subtilis). The replacement sporulation medium used is composed of inorganic salts and 10 mm ammonium lactate or glutamate. The requirement for ammonium lactate or glutamate could also be satisfied by other compounds that are metabolized via the tricarboxylic acid cycle. Sporulation of the suspended vegetative cells was completed by 8 to 10 hr after suspension, and the resulting spores were indistinguishable from spores produced in a conventional growth and sporulation medium. Various physiological changes previously reported to be associated with sporulation (e.g., increase in the level of tricarboxylic acid cycle enzymes and changes in the rates of synthesis of deoxyribonucleic acid, ribonucleic acid, and protein) could also be demonstrated during replacement sporulation.

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Selected References

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

  1. Anagnostopoulos C., Spizizen J. REQUIREMENTS FOR TRANSFORMATION IN BACILLUS SUBTILIS. J Bacteriol. 1961 May;81(5):741–746. doi: 10.1128/jb.81.5.741-746.1961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. BERNLOHR R. W., NOVELLI G. D. BACITRACIN BIOSYNTHESIS AND SPORE FORMATION: THE PHYSIOLOGICAL ROLE OF AN ANTIBIOTIC. Arch Biochem Biophys. 1963 Oct;103:94–104. doi: 10.1016/0003-9861(63)90014-6. [DOI] [PubMed] [Google Scholar]
  3. BLACK S. H., GERHARDT P. "Endotrophic" sporulation. Ann N Y Acad Sci. 1963 Jan 21;102:755–762. doi: 10.1111/j.1749-6632.1963.tb13674.x. [DOI] [PubMed] [Google Scholar]
  4. Bernlohr R. W. Changes in amino acid permeation during sporulation. J Bacteriol. 1967 Mar;93(3):1031–1044. doi: 10.1128/jb.93.3.1031-1044.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Boylen C. W., Ensign J. C. Ratio of teichoic acid and peptidoglycan in cell walls of Bacillus subtilis following spire germination and during vegetative growth. J Bacteriol. 1968 Aug;96(2):421–427. doi: 10.1128/jb.96.2.421-427.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. CANFIELD R. E., SZULMAJSTER J. TIME OF SYNTHESIS OF DEOXYRIBONUCLEIC ACID AND PROTEIN IN SPORES OF B. SUBTILIS. Nature. 1964 Aug 8;203:596–598. doi: 10.1038/203596a0. [DOI] [PubMed] [Google Scholar]
  7. DOI R. H., IGARASHI R. T. GENETIC TRANSCRIPTION DURING MORPHOGENESIS. Proc Natl Acad Sci U S A. 1964 Sep;52:755–762. doi: 10.1073/pnas.52.3.755. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. DONNELLAN J. E., Jr, NAGS E. H., LEVINSON H. S. CHEMICALLY DEFINED, SYNTHETIC MEDIA FOR SPORULATION AND FOR GERMINATION AND GROWTH OF BACILLUS SUBTILIS. J Bacteriol. 1964 Feb;87:332–336. doi: 10.1128/jb.87.2.332-336.1964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Eberle H., Lark K. G. Chromosome replication in Bacillus subtilis cultures growing at different rates. Proc Natl Acad Sci U S A. 1967 Jan;57(1):95–101. doi: 10.1073/pnas.57.1.95. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. FREESE E., PARK S. W., CASHEL M. THE DEVELOPMENTAL SIGNIFICANCE OF ALANINE DEHYDROGENASE IN BACILLUS SUBTILIS. Proc Natl Acad Sci U S A. 1964 Jun;51:1164–1172. doi: 10.1073/pnas.51.6.1164. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. HANSON R. S., SRINIVASAN V. R., HALVORSON H. O. BIOCHEMISTRY OF SPORULATION. II. ENZYMATIC CHANGES DURING SPORULATION OF BACILLUS CEREUS. J Bacteriol. 1963 Jul;86:45–50. doi: 10.1128/jb.86.1.45-50.1963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hanson R. S., Cox D. P. Effect of different nutritional conditions on the synthesis of tricarboxylic acid cycle enzymes. J Bacteriol. 1967 Jun;93(6):1777–1787. doi: 10.1128/jb.93.6.1777-1787.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Holmes P. K., Levinson H. S. Metabolic requirements for microcycle sporogenesis of Bacillus megaterium. J Bacteriol. 1967 Aug;94(2):434–440. doi: 10.1128/jb.94.2.434-440.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. MONRO R. E. Protein turnover and the formation of protein inclusions during sporulation of Bacillus thuringiensis. Biochem J. 1961 Nov;81:225–232. doi: 10.1042/bj0810225. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. MacKechnie I., Hanson R. S. Microcycle sporogenesis of Bacillus cereus in a chemically defined medium. J Bacteriol. 1968 Feb;95(2):355–359. doi: 10.1128/jb.95.2.355-359.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Mandelstam J., Waites W. M. Sporulation in Bacillus subtilis. The role of exoprotease. Biochem J. 1968 Oct;109(5):793–801. doi: 10.1042/bj1090793. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. NAKATA H. M., HALVORSON H. O. Biochemical changes occurring during growth and sporulation of Bacillus cereus. J Bacteriol. 1960 Dec;80:801–810. doi: 10.1128/jb.80.6.801-810.1960. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. POWELL J. F. Factors affecting the germination of thick suspensions of bacillus subtilis spores in L-alanine solution. J Gen Microbiol. 1950 Sep;4(3):330–338. doi: 10.1099/00221287-4-3-330. [DOI] [PubMed] [Google Scholar]
  20. RAMALEY R. F., BERNLOHR R. W. APPARENT INDUCTION OF ORNITHINE TRANSCARBAMYLASE AND ARGINASE BY ARGININE IN BACILLUS LICHENIFORMIS. J Mol Biol. 1965 Apr;11:842–844. doi: 10.1016/s0022-2836(65)80041-9. [DOI] [PubMed] [Google Scholar]
  21. SACKS L. E., ALDERTON G. Behavior of bacterial spores in aqueous polymer two-phase systems. J Bacteriol. 1961 Sep;82:331–341. doi: 10.1128/jb.82.3.331-341.1961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. SZULMAJSTER J. BIOCHIMIE DE LA SPOROG'EN'ESE CHEZ B. SUBTILIS. Bull Soc Chim Biol (Paris) 1964;46:443–481. [PubMed] [Google Scholar]
  23. TAKAHASHI I. LOCALIZATION OF SPORE MARKERS ON THE CHROMOSOME OF BACILLUS SUBTILIS. J Bacteriol. 1965 Apr;89:1065–1067. doi: 10.1128/jb.89.4.1065-1067.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Tamir H., Gilvarg C. Density gradient centrifugation for the separation of sporulating forms of bacteria. J Biol Chem. 1966 Mar 10;241(5):1085–1090. [PubMed] [Google Scholar]
  25. Vinter V., Slepecky R. A. Direct Transition of Outgrowing Bacterial Spores to New Sporangia Without Intermediate Cell Division. J Bacteriol. 1965 Sep;90(3):803–807. doi: 10.1128/jb.90.3.803-807.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Warren S. C. Sporulation in Bacillus subtilis. Biochemical changes. Biochem J. 1968 Oct;109(5):811–818. doi: 10.1042/bj1090811. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Wax R., Freese E., Cashel M. Separation of two functional roles of L-alanine in the initiation of Bacillus subtilis spore germination. J Bacteriol. 1967 Sep;94(3):522–529. doi: 10.1128/jb.94.3.522-529.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]

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