Abstract
Dipicolinic acid (pyridine-2,6-carboxylic acid; DPA) is a major component of bacterial spores and has been shown to be an important determinant of spore resistance. In the core of dormant Bacillus subtilis spores, DPA is associated with divalent calcium in a 1:1 chelate (Ca–DPA). Spores excrete Ca–DPA during germination, but it is unknown whether Ca and DPA are imported separately or together into the developing spore. Elemental analysis by scanning electron microscopy–energy-dispersive X-ray spectroscopy (SEM–EDS) of wild-type spores and mutant spores lacking the ability to synthesize DPA showed that DPA-less spores also lacked calcium, suggesting that the two compounds may be co-imported.
This is a preview of subscription content, log in via an institution to check access.
References
Daniel RA, Errington J (1993) Cloning, DNA sequence, functional analysis and transcriptional regulation of the genes encoding dipicolinic acid synthetase required for sporulation in Bacillus subtilis. J Mol Biol 232:468–483
Fort P, Errington J (1985) Nucleotide sequence and complementation analysis of a polycistronic sporulation operon, spoVA, in Bacillus subtilis. J Gen Microbiol 131:1091–1105
Hanson RS, Curry MV, Garner JV, Halvorson HO (1972) Mutants of Bacillus cereus strain T that produce thermoresistant spores lacking dipicolinate and have low levels of calcium. Can J Microbiol 18:1139–1143
Huang S, Chen D, Pelczar PL, Vepachedu VR, Setlow P, Li Y (2007) Levels of Ca2+-dipicolinic acid in individual Bacillus spores determined using microfluidic Raman tweezers. J Bacteriol 189:4681–4687
Miller J (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY
Moir A, Smith DA (1990) The genetics of bacterial spore germination. Annu Rev Microbiol 44:531–553
Moir A, Lafferty E, Smith DA (1979) Genetic analysis of spore germination mutants of Bacillus subtilis 168: the correlation of phenotype and map location. J Gen Microbiol 111:165–180
Murrell WG (1967) The biochemistry of the bacterial spore. Adv Microb Physiol 1:133–251
Murrell WG, Warth AG (1965) Composition and heat resistance of bacterial spores. In: Campbell LL, Halvorson HO (eds) Spores III. American Society for Microbiology, Washington DC, pp 1–24
Nicholson WL (2002) Roles of Bacillus spores in the environment. Cell Mol Life Sci 59:410–416
Nicholson WL (2004) Ubiquity, longevity, and ecological roles of Bacillus spores. In: Ricca E, Henriques AO, Cutting SM (eds) Bacterial spore formers: probiotics and emerging applications. Horizon Scientific Press, Norfolk, pp 1–15
Nicholson WL, Setlow P (1990) Chapter 9. Sporulation, germination, and outgrowth. In: Harwood CR, Cutting SM (eds) Molecular biological methods for Bacillus. Wiley, New York, pp 391–450
Nicholson WL, Munakata N, Horneck G, Melosh HJ, Setlow P (2000) Resistance of bacterial endospores to extreme terrestrial and extraterrestrial environments. Microbiol Mol Biol Rev 64:548–572
Paidhungat M, Setlow P (1999) Isolation and characterization of mutations in Bacillus subtilis that allow spore germination in the novel germinant d-alanine. J Bacteriol 181:3341–3350
Paidhungat M, Setlow P (2000) Role of Ger proteins in nutrient and nonnutrient triggering of spore germination in Bacillus subtilis. J Bacteriol 182:2513–2519
Paidhungat M, Setlow B, Driks A, Setlow P (2000) Characterization of spores of Bacillus subtilis which lack dipicolinic acid. J Bacteriol 182:5505–5512
Peng L, Chen D, Setlow P, Li Y (2009) Elastic and inelastic light scattering from single bacterial spores in an optical trap allows the monitoring of spore germination dynamics. Anal Chem 81:4035–4042
Rotman Y, Fields ML (1967) A modified reagent for dipicolinic acid analysis. Anal Biochem 22:168
Sammons RL, Moir A, Smith DA (1981) Isolation and properties of spore germination mutants of Bacillus subtilis 168 deficient in the initiation of germination. J Gen Microbiol 124:229–241
Schaeffer P, Millet J, Aubert J-P (1965) Catabolic repression of bacterial sporulation. Proc Natl Acad Sci USA 54:704–711
Setlow P (2006) Spores of Bacillus subtilis: their resistance to and killing by radiation, heat, and chemicals. J Appl Microbiol 101:514–525
Setlow B, Setlow P (1993) Dipicolinic acid greatly enhances production of spore photoproduct in bacterial spores upon UV irradiation. Appl Environ Microbiol 59:640–643
Slieman TA, Nicholson WL (2001) Role of dipicolinic acid in resistance of Bacillus subtilis spores exposed to artificial and solar UV radiation. Appl Environ Microbiol 67:1274–1279
Stewart M, Somylo AP, Somlyo AV, Shuman H, Lindsay JA, Murrell WG (1980) Distribution of calcium and other elements in cryosectioned Bacillus cereus T spores, determined by high-resolution scanning electron probe X-ray analysis. J Bacteriol 143:481–491
Tovar-Rojo F, Chander M, Setlow B, Setlow P (2002) The products of the spoVA operon are involved in dipicolinic acid uptake into developing spores of Bacillus subtilis. J Bacteriol 184:584–587
Acknowledgments
The authors wish to thank Pete Setlow for generous donation of the strains used. This work was supported by USDA grant FLA-MCS-04602 to W.L.N.
Author information
Authors and Affiliations
Corresponding author
Additional information
Communicated by Erko Stackebrandt.
Rights and permissions
About this article
Cite this article
Hintze, P.E., Nicholson, W.L. Single-spore elemental analyses indicate that dipicolinic acid-deficient Bacillus subtilis spores fail to accumulate calcium. Arch Microbiol 192, 493–497 (2010). https://doi.org/10.1007/s00203-010-0569-5
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00203-010-0569-5