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Applied and Environmental Microbiology, December 2007, p. 7747-7752, Vol. 73, No. 23
0099-2240/07/$08.00+0     doi:10.1128/AEM.00757-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Analysis of Nitrogen-Fixing Members of the {varepsilon} Subclass of Proteobacteria in Salt Marsh Sediments{triangledown}

Allana Welsh,1 David J. Burke,2 and Dittmar Hahn1*

Department of Biology, Texas State University, 601 University Drive, San Marcos, Texas 78666,1 The Holden Arboretum, 9500 Sperry Road, Kirtland, Ohio 4409422

Received 4 April 2007/ Accepted 26 September 2007

Based on phylogenetic analysis of clones retrieved from two nifH gene clone libraries that were created using cDNA from suboxic sediment samples obtained from areas densely vegetated with the high-salt marsh plant Spartina patens, a primer set was designed to target nitrogen-fixing bacteria with sequence similarities to members of the {varepsilon} subclass of Proteobacteria. Nested PCR, denaturing gel electrophoresis, and subsequent sequence analysis of reamplified fragments confirmed the specificity of the primer set by retrieving nifH sequences of only putative members of the {varepsilon} subclass of Proteobacteria, all of which were characterized by a highly divergent 27- or 36-bp insertion in both DNA and cDNA.

* Corresponding author. Mailing address: Department of Biology, Texas State University, 601 University Drive, San Marcos, TX 78666. Phone: (512) 245-3372. Fax: (512) 245-8713. E-mail: dh49{at}txstate.edu

{triangledown} Published ahead of print on 5 October 2007.

Applied and Environmental Microbiology, December 2007, p. 7747-7752, Vol. 73, No. 23
0099-2240/07/$08.00+0     doi:10.1128/AEM.00757-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.





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