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Applied and Environmental Microbiology, September 2000, p. 3814-3822, Vol. 66, No. 9
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Molecular Analysis of Diazotroph Diversity in the Rhizosphere of the Smooth Cordgrass, Spartina alterniflora

Charles R. Lovell,* Yvette M. Piceno,dagger Joseph M. Quattro, and Christopher E. Bagwell

Department of Biological Sciences, University of South Carolina, Columbia, South Carolina 29208

Received 22 March 2000/Accepted 11 July 2000

N2 fixation by diazotrophic bacteria associated with the roots of the smooth cordgrass, Spartina alterniflora, is an important source of new nitrogen in many salt marsh ecosystems. However, the diversity and phylogenetic affiliations of these rhizosphere diazotrophs are unknown. Denaturing gradient gel electrophoresis (DGGE) of PCR-amplified nifH sequence segments was used in previous studies to examine the stability and dynamics of the Spartina rhizosphere diazotroph assemblages in the North Inlet salt marsh, near Georgetown, S.C. In this study, plugs were taken from gel bands from representative DGGE gels, the nifH amplimers were recovered and cloned, and their sequences were determined. A total of 59 sequences were recovered, and the amino acid sequences predicted from them were aligned with sequences from known and unknown diazotrophs in order to determine the types of organisms present in the Spartina rhizosphere. We recovered numerous sequences from diazotrophs in the gamma  subdivision of the division Proteobacteria (gamma -Proteobacteria) and from various anaerobic diazotrophs. Diazotrophs in the alpha -Proteobacteria were poorly represented. None of the Spartina rhizosphere DGGE band sequences were identical to any known or previously recovered environmental nifH sequences. The Spartina rhizosphere diazotroph assemblage is very diverse and apparently consists mainly of unknown organisms.

* Corresponding author. Mailing address: Department of Biological Sciences, University of South Carolina, Columbia, SC 29208. Phone: (803) 777-7036. Fax: (803) 777-4002. E-mail: lovell{at}biol.sc.edu.

dagger Present address: Microbial Insights, Inc., Rockford, TN 37853-3044.

Applied and Environmental Microbiology, September 2000, p. 3814-3822, Vol. 66, No. 9
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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