Archives of Microbiology

, Volume 171, Issue 4, pp 265–272

Specific detection of green sulfur bacteria by in situ hybridization with a fluorescently labeled oligonucleotide probe

  • Christian Tuschak
  • Jens Glaeser
  • J. Overmann
Original paper

DOI: 10.1007/s002030050709

Cite this article as:
Tuschak, C., Glaeser, J. & Overmann, J. Arch Microbiol (1999) 171: 265. doi:10.1007/s002030050709
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Abstract

An oligodeoxynucleotide probe (GSB-532) specific for green sulfur bacteria was developed. Highly stringent hybridization conditions were established using whole cells of Chlorobium limicola DSM249 immobilized on glass slides. At a formamide concentration of 10%, the optimum specificity was reached at 47 °C. When a conventional fixation procedure was used, a conspicuous autofluorescence developed within the cells. This autofluorescence was due to the liberation of bacteriochlorophyll by the detergent Triton X-100 and a subsequent conversion to bacteriopheophytin and related compounds. The signal-to-noise ratio could be increased by a final dehydration of the samples with methanol. Finally, the method was adapted to the hybridization of natural samples collected on polycarbonate membrane filters. In situ hybridization of pure cultures, various enrichments, and natural samples from the chemocline of a freshwater lake confirmed that probe GSB-532 hybridized exclusively to cells of green sulfur bacteria. Our protocol allows the highly specific detection of green sulfur bacteria in water samples and a rapid screening of natural bacterial communities. Employing probe GSB-532, the phylogenetic affiliation of the epibionts in “Chlorochromatium aggregatum” and “Pelochromatium roseum” could be demonstrated for the first time.

Key words Green sulfur bacteria Chlorobiaceae In situ hybridization Oligonucleotide probe “Chlorochromatium” “Pelochromatium” 

Copyright information

© Springer-Verlag Berlin Heidelberg 1999

Authors and Affiliations

  • Christian Tuschak
    • 1
  • Jens Glaeser
    • 1
  • J. Overmann
    • 1
  1. 1.Paläomikrobiologie, Institut für Chemie und Biologie des Meeres, Universität Oldenburg, Postfach 2503, D-26111 Oldenburg, Germany e-mail: j.overmann@palmikro.icbm.uni-oldenburg.de Tel. +49-441-798-5376; Fax +49-441-798-3583DE

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