This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Bender, K. S.
Right arrow Articles by Achenbach, L. A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Bender, K. S.
Right arrow Articles by Achenbach, L. A.
Agricola
Right arrow Articles by Bender, K. S.
Right arrow Articles by Achenbach, L. A.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, September 2004, p. 5651-5658, Vol. 70, No. 9
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.9.5651-5658.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Metabolic Primers for Detection of (Per)chlorate-Reducing Bacteria in the Environment and Phylogenetic Analysis of cld Gene Sequences

Kelly S. Bender,1,{dagger} Melissa R. Rice,1 William H. Fugate,1 John D. Coates,2 and Laurie A. Achenbach1*

Department of Microbiology, Southern Illinois University, Carbondale, Illinois,1 Department of Plant and Microbial Biology, University of California Berkeley, Berkeley, California2

Received 26 April 2004/ Accepted 22 May 2004

Natural attenuation of the environmental contaminant perchlorate is a cost-effective alternative to current removal methods. The success of natural perchlorate remediation is dependent on the presence and activity of dissimilatory (per)chlorate-reducing bacteria (DPRB) within a target site. To detect DPRB in the environment, two degenerate primer sets targeting the chlorite dismutase (cld) gene were developed and optimized. A nested PCR approach was used in conjunction with these primer sets to increase the sensitivity of the molecular detection method. Screening of environmental samples indicated that all products amplified by this method were cld gene sequences. These sequences were obtained from pristine sites as well as contaminated sites from which DPRB were isolated. More than one cld phylotype was also identified from some samples, indicating the presence of more than one DPRB strain at those sites. The use of these primer sets represents a direct and sensitive molecular method for the qualitative detection of (per)chlorate-reducing bacteria in the environment, thus offering another tool for monitoring natural attenuation. Sequences of cld genes isolated in the course of this project were also generated from various DPRB and provided the first opportunity for a phylogenetic treatment of this metabolic gene. Comparisons of the cld and 16S ribosomal DNA (rDNA) gene trees indicated that the cld gene does not track 16S rDNA phylogeny, further implicating the possible role of horizontal transfer in the evolution of (per)chlorate respiration.

* Corresponding author. Mailing address: Department of Microbiology, Southern Illinois University, Carbondale, IL 62901. Phone: (618) 453-7984. Fax: (618) 453-8036. E-mail: laurie{at}micro.siu.edu.

{dagger} Present address: Department of Biochemistry, University of Missouri—Columbia, Columbia, Mo.

Applied and Environmental Microbiology, September 2004, p. 5651-5658, Vol. 70, No. 9
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.9.5651-5658.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Weelink, S. A. B., Tan, N. C. G., ten Broeke, H., van den Kieboom, C., van Doesburg, W., Langenhoff, A. A. M., Gerritse, J., Junca, H., Stams, A. J. M. (2008). Isolation and Characterization of Alicycliphilus denitrificans Strain BC, Which Grows on Benzene with Chlorate as the Electron Acceptor. Appl. Environ. Microbiol. 74: 6672-6681 [Abstract] [Full Text]  
  • Lee, A. Q., Streit, B. R., Zdilla, M. J., Abu-Omar, M. M., DuBois, J. L. (2008). Mechanism of and exquisite selectivity for O-O bond formation by the heme-dependent chlorite dismutase. Proc. Natl. Acad. Sci. USA 105: 15654-15659 [Abstract] [Full Text]  
  • Nozawa-Inoue, M., Jien, M., Hamilton, N. S., Stewart, V., Scow, K. M., Hristova, K. R. (2008). Quantitative Detection of Perchlorate-Reducing Bacteria by Real-Time PCR Targeting the Perchlorate Reductase Gene. Appl. Environ. Microbiol. 74: 1941-1944 [Abstract] [Full Text]  
  • Lalucat, J., Bennasar, A., Bosch, R., Garcia-Valdes, E., Palleroni, N. J. (2006). Biology of Pseudomonas stutzeri. Microbiol. Mol. Biol. Rev. 70: 510-547 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»