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Thymidine incorporation of bacteria sequentially extracted from soil using repeated homogenization-centrifugation

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Abstract

Bacteria were sequentially extracted from soil into a water suspension after shaking soil with water or mixing it in a blender followed by a low-speed centrifugation. Bacteria, which were released only after several cycles of homogenization-centrifugation, had higher growth rates as judged from thymidine and leucine incorporation, whereas bacteria that were more readily released by a gentle shaking procedure had the lowest growth rate. This indicated that bacteria more tightly bound to soil particles were growing faster than those that were more easily released into the water suspension. The same pattern was found both in an agricultural and a forest soil, with contrasting pH and organic matter content, and irrespective of whether the bacteria were labeled before or after the centrifugation steps. The different growth rates of the bacteria could not be explained by different partitioning of label between different macromolecules, different cell size, different viability of the bacteria, or different dilution of the added radioactive substrate in the different homogenization-centrifugation fractions. The total amount of phospholipid fatty acids per bacterial cell was also similar in the different fractions. Different composition of the bacterial communities in the different homogenization-centrifugation fractions was indicated by a gradually altered phospholipid fatty acid pattern of the extracted bacteria, and an increased hydrophobicity of the bacteria released only after several homogenization-centrifugation treatments.

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References

  1. Bååth E (1992) Measurement of heavy metal tolerance of soil bacteria using thymidine incorporation into bacteria extracted after homogenization-centrifugation. Soil Biol Biochem 24:1167–1172

    Article  Google Scholar 

  2. Bååth E (1992) Thymidine incorporation into macromolecules of bacteria extracted from soil by homogenization-centrifugation. Soil Biol Biochem 24:1157–1165

    Article  Google Scholar 

  3. Bååth E (1994) Measurement of protein synthesis of soil bacteria assemblages using the leucine incorporation technique. Biol Fertil Soils 17:147–153

    Article  Google Scholar 

  4. Bååth E (1994) Thymidine and leucine incorporation in soil bacteria with different cell size. Microb Ecol 27:267–278

    Article  Google Scholar 

  5. Bakken LR (1985) Separation and purification of soil bacteria by density gradient centrifugation. Appl Environ Microbiol 49:1482–1487

    Article  CAS  Google Scholar 

  6. Bakken LR, Olsen RA (1987) The relationship between cell size and viability of soil bacteria. Microb Ecol 13:103–114

    Article  CAS  Google Scholar 

  7. Bakken LR, Olsen RA (1989) DNA-content of soil bacteria of different cell size. Soil Biol Biochem 21:789–793

    Article  Google Scholar 

  8. Christensen H (1993) Conversion factors for the thymidine incorporation technique estimated with bacteria in pure culture and on seedling roots. Soil Biol Biochem 25:1085–1096

    Article  CAS  Google Scholar 

  9. Cortez J, Schnitzer M (1979) Nucleic acid bases in soils and their association with organic and inorganic soil components. Can J Soil Sci 59:277–286

    Article  CAS  Google Scholar 

  10. Díaz-Raviña M, Bååth E, Frostegåd A (1994) Multiple heavy metal tolerance of soil bacterial communities and its measurement by a thymidine incorporation technique. Appl Environ Microbiol 60:2238–2247

    Article  Google Scholar 

  11. Díaz-Raviña M, Frostegård A, Bååth E. (1994) Thymidine, leucine and acetate incorporation into soil bacterial assemblages at different temperatures. FEMS Microbiol Ecol 14:221–232

    Article  Google Scholar 

  12. Faegri A, Lid Torsvik V, Goksöyr J (1977) Bacterial and fungal activities in soil: separation of bacteria and fungi by a rapid fractionated centrifugation technique. Soil Biol Biochem 9:105–112

    Article  Google Scholar 

  13. Frostegård Å, Bååth E (1996) The use of phospholipid fatty acid analysis to estimate bacterial and fungal biomass in soil. Biol Fert Soils in press

  14. Frostegård Å, Tunlid A, Bååth E (1993) Phospholipid fatty acid composition, biomass and activity of microbial communities from two soil types exposed to different heavy metals. Appl Environ Microbiol 59:3605–3617

    Article  Google Scholar 

  15. Hahn D, Kester R, Starrenburg MJC, Akkermans ADL (1990) Extraction of ribosomal RNA from soil for detection of Frankia with oligonucleotide probes. Arch Mikrobiol 154:329–335

    Article  CAS  Google Scholar 

  16. Hattori T (1982) Analysis of plate count data of bacteria in natural environments. J Gen Appl Microbiol 28:13–22

    Article  Google Scholar 

  17. HolbenWE, Jansson JK, Chelm BK, Tiedje JM (1988) DNA probe method for the detection of specific microorganisms in the soil bacterial community. Appl Environ Microbiol 54:703–711

    Article  CAS  Google Scholar 

  18. Hopkins DW Macnaughton SJ, O'Donell AG (1991) A dispersion and differential centrifugation technique for representatively sampling microorganisms from soil. Soil Biol Biochem 23:217–225

    Article  Google Scholar 

  19. Hopkins DW O'Donell AG, Macnaughton SJ (1991) Evaluation of a dispersion and elutriation technique for sampling microorganisms from soil. Soil Biol Biochem 23:227–232

    Article  Google Scholar 

  20. Huysman F, Verstraete W (1993) Effect of cell surface characteristics on the adhesion of bacteria to soil particles. Biol Fertil Soils 16:21–26

    Article  Google Scholar 

  21. Huysman F, Verstraete W (1993) Water-facilitated transport of bacteria in unsaturated soil columns: influence of cell surface hydrophobicity and soil properties. Soil Biol Biochem 25:83–90

    Article  Google Scholar 

  22. Jacobsen CS, Rasmussen OF (1992) Development and application of a new method to extract bacterial DNA from soil based on separation of bacteria from soil with cation-exchange resin. Appl Environ Microbiol 58:2458–2462

    Article  CAS  Google Scholar 

  23. Kasahara Y, Morisaki H, Hattori T (1993) Hydrophobicity of the cells of fast- and slow-growing bacteria isolated from a grassland soil. J Gen Appl Microbiol 39:381–388

    Article  CAS  Google Scholar 

  24. Krambeck C, Krambeck H-J, Overbeck J (1981) Microcomputer assisted biomass determination of plankton bacteria on scanning electron micrographs. Appl Environ Microbiol 42:142–149

    Article  CAS  Google Scholar 

  25. Macdonald RM (1986) Sampling soil microfloras: dispersion of soil by ion exchange and extraction of specific microorganisms by elutriation. Soil Biol Biochem 18:399–406

    Article  Google Scholar 

  26. Macdonald RM (1986) Sampling soil microfloras: Optimization of density gradient centrifugation in Percoll to separate microorganisms from soil suspensions. Soil Biol Biochem 18:407–410

    Article  Google Scholar 

  27. Pedrós-Alió P, Brock TD (1983) The importance of attachment to particles for planctonic bacteria. Arch Hydrobiol 98:354–379

    Google Scholar 

  28. Pillai SD, Josephson KL, Bailey RL, Gerba CP, Pepper IL (1991) Rapid method for processing soil samples for polymerase chain reaction amplification of specific gene sequences. Appl Environ Microbiol 57:2283–2286

    Article  CAS  Google Scholar 

  29. Pollard PC, Moriarty DJW (1984) Validity of the tritiated thymidine method for estimating bacterial growth rates: measurement of isotope dilution during DNA synthesis. Appl Environ Microbiol 48:1076–1083

    Article  CAS  Google Scholar 

  30. Riemann B, Söndergaard M (1984) Measurement of diel rates of bacterial secondary production in aquatic environments. Appl Environ Microbiol 47:632–638

    Article  CAS  Google Scholar 

  31. Robarts RD, Zohary T (1993) Fact or fiction—Bacterial growth rates and production as determined by [methyl-3H]-thymidine? Adv Microb Ecol 13:371–425

    Article  CAS  Google Scholar 

  32. Simon M (1985) Specific uptake rates of aminoacids by attached and free-living bacteria in a mesotrophic lake. Appl Environ Microbiol 49:1254–1259

    Article  CAS  Google Scholar 

  33. Steffan RJ, Goksöyr J, Bej AK, Atlas RM (1988) Recovery of DNA from soils and sediments. Appl Environ Microbiol 54:2908–2915

    Article  CAS  Google Scholar 

  34. Torsvik VL (1980) Isolation of bacterial DNA from soil. Soil Biol Biochem 12:15–21

    Article  CAS  Google Scholar 

  35. Unanue M, Ayo B, Azúa I, Barcina I, Iriberri J (1992) Temporal variability of attached and freeliving bacteria in coastal waters. Microb Ecol 23:27–39

    Article  CAS  Google Scholar 

  36. van Loosdrecht CM, Lyklema J, Norde W Zehnder AJB (1990) Influence of interfaces on microbial activity. Microbiol Rev 54:75–87

    Article  Google Scholar 

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  1. Department of Microbial Ecology, Lund University, Ecology Building, S-223 62, Lund, Sweden

    Erland Bååth

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  1. Erland Bååth
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Correspondence to: E. Bååth

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Bååth, E. Thymidine incorporation of bacteria sequentially extracted from soil using repeated homogenization-centrifugation. Microb Ecol 31, 153–166 (1996). https://doi.org/10.1007/BF00167861

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