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Seasonal Variability of Diazotroph Assemblages Associated with the Rhizosphere of the Salt Marsh Cordgrass, Spartina alterniflora

  • Environmental Microbiology
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Abstract

Nitrogen fixation is the primary N source in the highly productive but N-limited North Inlet, SC, USA salt marsh system. The diverse assemblages of nitrogen-fixing (diazotrophic) bacteria associated with the rhizospheres of the short and tall growth forms of Spartina alterniflora were analyzed at two sites, Crab Haul Creek and Goat Island, which are in different tidal creek drainage systems in this marsh. The sites differed in proximity to the main channel for tidal intrusion and in several edaphic parameters. We hypothesized that either the differing abiotic environmental regimes of the two sites or the variation due to seasonal effects result in differences in the diazotroph assemblage. Rhizosphere samples were collected seasonally during 1999 and 2000. DNA was purified and nifH amplified for denaturing gradient gel electrophoresis (DGGE) analysis of diazotroph assemblage composition. Principal components analysis was used to analyze the binary DGGE band position data. Season strongly influenced assemblage composition and biplots were used to identify bands that significantly affected the seasonal and site-specific clustering. The types of organisms that were most responsive to seasonal or site variability were identified on the basis of DGGE band sequences. Seasonally responsive members of the anaerobic diazotrophs were detected during the winter and postsenescence conditions and may have been responsible for elevated pore water sulfide concentrations. Sequences from a diverse assemblage of Gammaproteobacteria were predominant during growth periods of S. alterniflora. Abiotic environmental parameters strongly influenced both the S. alterniflora and the diazotrophic bacterial assemblages associated with this keystone salt marsh plant species.

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Acknowledgements

We acknowledge the Belle W. Baruch Institute for Coastal and Marine Sciences for access to sampling sites and for logistical support and Mike Friez for assistance with DNA sequencing. This research was supported by the National Science Foundation awards DEB-0108412 and MCB-0237854 to C.R.L. Contribution 1578 to the Belle W. Baruch Institute for Coastal and Marine Sciences.

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Author notes

  1. Christopher E. Bagwell

    Present address: Environmental Sciences and Biotechnology, Savannah River National Laboratory, Aiken, SC, 29803, USA

  2. Jeannine LaRocque

    Present address: Slone-Kettering Institute, Memorial Slone-Kettering Cancer Center, New York, NY, 10065, USA

  3. Peter W. Bergholz

    Present address: Crop and Soil Sciences, Cornell University, Ithaca, NY, 14853, USA

Authors and Affiliations

  1. Department of Biological Sciences, University of South Carolina, 715 Sumter Street, Columbia, SC, 29208, USA

    Megan D. Gamble, Christopher E. Bagwell, Jeannine LaRocque, Peter W. Bergholz & Charles R. Lovell

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Correspondence to Charles R. Lovell.

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Supplementary Figure 1

Phylogenetic analysis of all bands from band stab analysis (nucleotide sequences, neighbor joining, 1,000 bootstrap replicates, Jukes Cantor correction and complete deletion of gaps and missing data) (GIF 1.27 mb)

High-resolution image (TIFF 8.91 mb)

Supplementary Figure 2

DGGE gel image representing September short- and tall-form Goat Island samples (GIF 621 kb)

High-resolution image (TIFF 300 kb)

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Gamble, M.D., Bagwell, C.E., LaRocque, J. et al. Seasonal Variability of Diazotroph Assemblages Associated with the Rhizosphere of the Salt Marsh Cordgrass, Spartina alterniflora . Microb Ecol 59, 253–265 (2010). https://doi.org/10.1007/s00248-009-9558-0

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