Abstract
This protocol defines conditions under which the germination of spores can be used to synchronize Bacillus subtilis cells, utilizing the time-ordered sequence of events taking place during the transition from spore to vegetative cells. The transition stages involve: phase change, swelling, emergence, initial division, and elongation. By using this method we have obtained two distinctive synchronized cell cycles, while the synchrony faded away in the third cycle. The advantage of using spore outgrowth and germination is that a highly synchronized population of bacterial cells can be obtained. The limitations of this method are that it can be applied only for sporulating bacteria and synchrony lasts for only a limited period of time exceeding not more than two cycles.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Graumann, P. (Ed.). (2007) Bacillus: Cellular and molecular biology (1st Ed.). Caister Academic Press, Wymondham. http://www.horizonpress.com/bac
Hansen, J. N., Spiegelman, G., and Halvorson, H. O. (1970) Bacterial spore outgrowth: Its regulation. Science 168, 1291–1298.
Keynan, A. (1973) The transformation of bacterial endospores into vegetative cells. Symp. Soc. Gen. Microbiol. 23, 85–123.
Mandelstam, J. (1976) Bacterial sporulation: A problem in the biochemistry and genetics of a primitive developmental system. Proc. R. Soc. B. 193, 89–106.
Nelson, D. L., and Kornberg, A. (1970) Biochemical studies of bacterial sporulation and germination. XVIII. Free amino acids in spores. J. Biol. Chem. 245, 1128–1136.
Setlow, P., and Kornberg, A. (1970) Biochemical studies of bacterial sporulation and germination. XXIII. Nucleotide metabolism during spore germination. J. Biol. Chem. 245, 3645–3652.
Balassa, G. (1965) Synthesis macromoleculaires au cours de la germination des spores de B. subtilis. I. Cinctique Annales De I’lnstitut Pasteur 109, 13–35.
Balassa, G. (1969) Biochemical genetics of bacterial sporulation. I. Unidirectional pleiotropic interactions among genes controlling sporulation in Bacillus subtilis. Mol. Gen. Genet. 104, 73–103.
Kobayashi, I, Steinberg, W., Higa, A., Halvorson, H. O., and Levinthal, C. (1965) Sequential synthesis of macromolecules during outgrowth of bacterial spores. In Spores IZZ, pp. 200–212. L. L. Campbell and H. O. Halvorson (Eds.). Washington, DC: American Society for Microbiology.
Torriani, A., and Levinthal, C. (1967) Ordered synthesis of proteins during outgrowth of spores of B. Cereus. J. Bacteriol. 94, 176–183.
Armstrong, R. L., and Sueoka, N. (1968) Phase transition in ribonucleic acid synthesis during germination of B. subtilis spores. Proc. Natl. Acad. Sci. USA 59, 153–160.
Steinberg, W., and Halvorson, H. O. (1968) Timing of enzyme synthesis during outgrowth of Bacillus Cereus. I. Ordered enzyme synthesis. J. Bacteriol. 95, 469–478.
Kennett, R. N., and Sueoka, N. (1971) Gene expression during outgrowth of B. subtilis spores. The relationship between order on the chromosome and temporal sequence of enzyme synthesis. J. Mol. Biol. 60, 31–44.
Campbell, L. L., Jr. (1957) In Spores, p. 33. H. Halvorson (Ed.). Washington, DC: American Institute of Biological Sciences.
Levinson, H. S., and Hyatt, M. T. (1956) Correlation of respiratory activity with phases of spore germination and growth in Bacillus megaterium as influenced by manganese and L-alanine. J. Bacteriol. 72, 176–183.
Halvorson, H. O. (1959) Symposium on initiation of bacterial growth. Bacteriol. Rev. 23, 267–272.
Harrell, W. K., and Halvorson, H. (1955) Studies on the role of L-alanine in the germination of spores of Bacillus terminalis. J. Bacteriol. 69, 275–279.
Yasuda, Y., and Tochikubo, K. (1984) Relation between D-glucose and L- and D-alanine in the initiation of germination of Bacillus subtilis spores. Microbiol. Immunol. 28, 197–207.
McCann, K. P., Robinson, C., Sammons, R. L., Smith, D. A., and Corfe, B. M. (1996) Alanine germination receptors of Bacillus subtilis. Lett. Appl. Microbiol. 23, 290–294.
Liang, L., Gai, Y., Hu, K., and Liu, G. (2008) The gerA operon is required for spore germination in Bacillus thuringiensis. Wei Sheng Wu Xue Bao. 48, 281–286. (Article in Chinese)
Powell, J. F. (1950) Factors affecting the germination of thick suspensions of Bacillus subtilis spores in L-alanine solution. J. Gen. Microbiol. 4, 330–338.
Pulvertaft, R. J. V., and Haynes, J. A. (1951) Adenosine and spore germination; phasecontrast studies. J. Gen. Microbiol. 5, 657–663.
Powell, J. F., and Strange, R. E. (1953) Biochemical changes occurring during the germination of bacterial spores. Biochem. J. 54, 205–209.
Murrell, W. G., and Scott W. J. (1958) The permeability of bacterial spores to water. Proc. 7th Int. Congr. Microbiol. Stockholm, p. 26.
Powell, J. F. (1957) Biochemical changes occurring during spore germination in bacillus species. J. Appl. Bacteriol. 20, 349–358.
Bhattacharya, S., and Sarkar, N. (1981) Study of deoxyribonucleic acid replication in permeable cells of Bacillus subtilis using mercurated nucleotide substrates. Biochemistry 20, 3029–3034.
Woese, C., and Morowitz, H. J. (1958) Kinetics of the release of dipicolinic acid from spores of Bacillus subtilis. J. Bacteriol. 76, 81–83.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2011 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Banfalvi, G. (2011). Synchronization of Bacillus subtilis Cells by Spore Germination and Outgrowth. In: Banfalvi, G. (eds) Cell Cycle Synchronization. Methods in Molecular Biology, vol 761. Humana Press. https://doi.org/10.1007/978-1-61779-182-6_11
Download citation
DOI: https://doi.org/10.1007/978-1-61779-182-6_11
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-61779-181-9
Online ISBN: 978-1-61779-182-6
eBook Packages: Springer Protocols