Abstract
In a soil/sediment model system we have shown recently that a gram-positive bacterium with natural competence (Bacillus subtilis) can take up transforming DNA adsorbed to sand minerals. Here we examined whether also a naturally transformable soil bacterium of the gramnegative pseudomonad (Pseudomonas stutzeri) can be transformed by mineral-associated DNA. for these studies the transformation protocol of this species was further improved and characterized. The peak of competence during growth of P. stutzeri was determined to occur at the beginning of the stationary phase. The competence state was conserved during shock freezing and thawing of cells in 10% glycerol. Kinetic experiments showed that transformant formation after addition of DNA to competent cells proceeded for more than 2 h with DNA adsorption to cells being the rate limiting step. By means of the defined protocol P. stutzeri was shown to be transformed by sand-adsorbed DNA. Transformation by adsorbed or dissolved DNA occurred between 16° and 44°C. Efficiency and DNaseI-sensitivity of transformation by DNA adsorbed to sand or in liquid were comparable. It is concluded that uptake of particle-bound DNA by P. stutzeri in soil is possible. This finding adds evidence to the view that transformation occurs in natural environments where DNA is assumed to be significantly associated with mineral/particulate material and thereby is protected against enzymatic degradation.
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Aardema BW, Lorenz MG, Krumbein WE (1983) Protection of sediment-adsorbed transforming DNA against enzymatic inactivation. Appl Environ Microbiol 46:417–420
Borenstein S, Ephrati-Elizur E (1969) Spontaneous release of DNA in sequential genetic order by Bacillus subtilis. J Mol Biol 45:137–152
Carlson CA, Pierson LS, Rosen JJ, Ingraham JL (1983) Pseudomonas stutzeri and related species undergo natural transformation. J Bacteriol 153:93–99
Catlin BW (1956) Extracellular deoxyribonucleic acid of bacteria and a deoxyribonuclease inhibitor. Science 124:441–442
Crabb DW, Streips UN, Doyle RJ (1977) Selective enrichment for genetic markers in DNA released by competent cultures of Bacillus subtilis. Mol Gen Genet 155: 179–183
Ephrati E (1968) Spontaneous transformation in Bacillus subtilis. Genet Res 11:83–96
Greaves MP, Wilson MJ (1970) The degradation of nucleic acids and montmorillonite-nucleic-acid complexes by soil microorganisms. Soil Biol Biochem 2:257–268
Hara T, Ueda S (1981) A study on the mechanism of DNA excretion from P. aeruginosa KYU-1—effect of mitomycin C on extracellular DNA production. Agric Biol Chem 45:2457–2461
Herdman M, Carr NG (1971) Recombination in Anacystis nidulans mediated by an extracellular DNA/RNA complex. J Gen Microbiol 68:XIV
Labarca CA, Paigen K (1980) A simple, rapid and sensitive DNA assay procedure. Anal Biochem 102:344–352
Lorenz MG, Wackernagel W (1987) Adsorption of DNA to sand and variable degradation rates of adsorbed DNA. Appl Environ Microbiol 53:2948–2952
Lorenz MG, Wackernagel W (1988) Impact of mineral surfaces on gene transfer by transformation in natural bacterial environments. In: Klingmüller W (ed) Risk assessment for deliberate releases. Springer, Berlin Heidelberg New York, p 110–119
Lorenz MG, Aardema BW, Krumbein WE (1981) Interaction of marine sediment with DNA and DNA avalability to nucleases. Mar Biol 64:225–230
Lorenz MG, Aardema BW, Wackernagel W (1988) Highly efficient genetic transformation of Bacillus subtilis attached to sand grains. J Gen Microbiol 134:107–112
Marmur J (1961) A procedure for the isolation of deoxyribonucleic acid from microorganisms. J Mol Biol 3:208–218
Ogram A, Sayler GS, Barkay T (1987) The extraction and purification of microbial DNA from sediments. J Microbiol Methods 7:57–66
Ogram A, Sayler GS, Gustin D, Lewis RJ (1988) DNA adsorption to soils and sediments. Environ Sci Technol 22:982–984
Sinha RP, Iyer UN (1971) Competence for genetic transformation and the release of DNA from Bacillus subtilis. Biochim Biophys Acta 232:61–71
Stewart GJ, Carlson CA (1986) The biology of natural transformation. Annu Rev Microbiol 40:211–235
Takahashi I (1962) Genetic transformation of Bacillus subtilis by extracellular DNA. Biochem Biophys Res Commun 6:467–470
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Lorenz, M.G., Wackernagel, W. Natural genetic transformation of Pseudomonas stutzeri by sand-adsorbed DNA. Arch. Microbiol. 154, 380–385 (1990). https://doi.org/10.1007/BF00276535
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DOI: https://doi.org/10.1007/BF00276535