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Applied and Environmental Microbiology, February 1999, p. 422-430, Vol. 65, No. 2
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Quantifying Microbial Diversity: Morphotypes, 16S rRNA Genes, and Carotenoids of Oxygenic Phototrophs in Microbial Mats

Ulrich Nübel, Ferran Garcia-Pichel,* Michael Kühl,dagger and Gerard MuyzerDagger

Max Planck Institute for Marine Microbiology, Bremen, Germany

Received 29 July 1998/Accepted 17 November 1998

We quantified the diversity of oxygenic phototrophic microorganisms present in eight hypersaline microbial mats on the basis of three cultivation-independent approaches. Morphological diversity was studied by microscopy. The diversity of carotenoids was examined by extraction from mat samples and high-pressure liquid chromatography analysis. The diversity of 16S rRNA genes from oxygenic phototrophic microorganisms was investigated by extraction of total DNA from mat samples, amplification of 16S rRNA gene segments from cyanobacteria and plastids of eukaryotic algae by phylum-specific PCR, and sequence-dependent separation of amplification products by denaturing-gradient gel electrophoresis. A numerical approach was introduced to correct for crowding the results of chromatographic and electrophoretic analyses. Diversity estimates typically varied up to twofold among mats. The congruence of richness estimates and Shannon-Weaver indices based on numbers and proportional abundances of unique morphotypes, 16S rRNA genes, and carotenoids unveiled the underlying diversity of oxygenic phototrophic microorganisms in the eight mat communities studied.

* Corresponding author. Mailing address: Max Planck Institute for Marine Microbiology, Celsiusstr. 1, D-28359 Bremen, Germany. Phone: 49-421-2028-838. Fax: 49-421-2028-580. E-mail: fgarcia{at}mpi-bremen.de.

dagger Present address: Marine Biological Laboratory, University of Copenhagen, 3000 Helsingør, Denmark.

Dagger Present address: Netherlands Institute for Sea Research (NIOZ), 1790 AB Den Burg (Texel), The Netherlands.

Applied and Environmental Microbiology, February 1999, p. 422-430, Vol. 65, No. 2
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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