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Applied and Environmental Microbiology, April 2001, p. 1893-1901, Vol. 67, No. 4
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.4.1893-1901.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Community Structure of Denitrifiers, Bacteria, and Archaea along Redox Gradients in Pacific Northwest Marine Sediments by Terminal Restriction Fragment Length Polymorphism Analysis of Amplified Nitrite Reductase (nirS) and 16S rRNA Genes

Gesche Braker,1,* Héctor L. Ayala-del-Río,1,2 Allan H. Devol,3 Andreas Fesefeldt,4 and James M. Tiedje1,2

Center for Microbial Ecology1 and Department of Microbiology and Molecular Genetics,2 Michigan State University, East Lansing, Michigan 48824-1325; School of Oceanography, University of Washington, Seattle, Washington 982953; and Landeskriminalamt Schleswig-Holstein, 24116 Kiel, Germany4

Received 17 August 2000/Accepted 19 December 2000

Steep vertical gradients of oxidants (O2 and NO3-) in Puget Sound and Washington continental margin sediments indicate that aerobic respiration and denitrification occur within the top few millimeters to centimeters. To systematically explore the underlying communities of denitrifiers, Bacteria, and Archaea along redox gradients at distant geographic locations, nitrite reductase (nirS) genes and bacterial and archaeal 16S rRNA genes (rDNAs) were PCR amplified and analyzed by terminal restriction fragment length polymorphism (T-RFLP) analysis. The suitablility of T-RFLP analysis for investigating communities of nirS-containing denitrifiers was established by the correspondence of dominant terminal restriction fragments (T-RFs) of nirS to computer-simulated T-RFs of nirS clones. These clones belonged to clusters II, III, and IV from the same cores and were analyzed in a previous study (G. Braker, J. Zhou, L. Wu, A. H. Devol, and J. M. Tiedje, Appl. Environ. Microbiol. 66:2096-2104, 2000). T-RFLP analysis of nirS and bacterial rDNA revealed a high level of functional and phylogenetic diversity, whereas the level of diversity of Archaea was lower. A comparison of T-RFLPs based on the presence or absence of T-RFs and correspondence analysis based on the frequencies and heights of T-RFs allowed us to group sediment samples according to the sampling location and thus clearly distinguish Puget Sound and the Washington margin populations. However, changes in community structure within sediment core sections during the transition from aerobic to anaerobic conditions were minor. Thus, within the top layers of marine sediments, redox gradients seem to result from the differential metabolic activities of populations of similar communities, probably through mixing by marine invertebrates rather than from the development of distinct communities.

* Corresponding author. Mailing address: Center for Microbial Ecology, 540 Plant and Soil Sciences Building, Michigan State University, East Lansing, MI 48824-1325. Phone: (517) 353-9021. Fax: (517) 353-2917. E-mail: braker{at}msu.edu.

Applied and Environmental Microbiology, April 2001, p. 1893-1901, Vol. 67, No. 4
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.4.1893-1901.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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