Analysis of Nitrogen-Fixing Members of the ε Subclass of Proteobacteria in Salt Marsh Sediments

• Published in: Applied and Environmental Microbiology Vol. 73; no. 23; pp. 7747 - 7752
• Main Authors: Welsh, Allana, Burke, David J, Hahn, Dittmar
• Format: Journal Article
• Language: English
• Published: Washington, DC American Society for Microbiology 01.12.2007
• Subjects: Animal, plant and microbial ecology; Bacterial Proteins - genetics; Biological and medical sciences; Brackish; DNA, Complementary - chemistry; DNA, Complementary - genetics; Epsilonproteobacteria - classification; Epsilonproteobacteria - genetics; Fundamental and applied biological sciences. Psychology; Geologic Sediments - microbiology; Microbial ecology; Microbiology; Molecular Sequence Data; Nitrogen Fixation - genetics; Oxidoreductases - genetics; Phylogeny; Proteobacteria; Sequence Analysis, DNA; Spartina patens; Various environments (extraatmospheric space, air, water); Wetlands; Nitrogen fixation; Salt marsh; Bacteria; Sediments
• ISSN: 0099-2240; 1098-5336
• DOI: 10.1128/AEM.00757-07

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 ε 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 ε subclass of Proteobacteria, all of which were characterized by a highly divergent 27- or 36-bp insertion in both DNA and cDNA.