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 Borneman, J.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Borneman, J.
Agricola
Right arrow Articles by Borneman, J.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, August 1999, p. 3398-3400, Vol. 65, No. 8
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Culture-Independent Identification of Microorganisms That Respond to Specified Stimuli

James Borneman*

Department of Plant Pathology, University of California, Riverside, California 92521

Received 5 February 1999/Accepted 17 May 1999

A new approach that permits culture-independent identification of microorganisms that respond to specified stimuli was developed. This approach was illustrated by examination of microorganisms that grew in response to various nutrient supplements added to soil. A thymidine nucleotide analog, bromodeoxyuridine (BrdU), and supplements were added to soil and incubated for 3 days. DNA was extracted from the soil, and the newly synthesized DNA was isolated by immunocapture of the BrdU-labeled DNA. The unique perspective this approach offers was demonstrated by comparing the microbial community structures obtained from total soil DNA and the BrdU-labeled fraction in an rRNA gene (rDNA) analysis. The traditional total DNA analysis revealed no notable differences between the treatments, whereas the BrdU-labeled DNA showed significantly different banding patterns between the nutrient supplement treatments and compared with total DNA banding patterns. PCR primers were developed to specifically amplify the intergenic region of an rDNA sequence unique to the BrdU analysis of a phosphate supplement treatment. Amplification of DNA from all treatments using these primers showed that it was unique to the phosphate treatment and that it was present in both the total DNA and BrdU-labeled DNA fractions. This result demonstrates the promise of this new strategy, because it was able to permit identification of a sequence from a phosphate-responsive organism that was not discernable in the traditional total DNA community structure analysis.

* Mailing address: Department of Plant Pathology, University of California, Riverside, CA 92521. Phone: 909-787-3584. Fax: 909-787-3782. E-mail: borneman{at}ucrac1.ucr.edu.

Applied and Environmental Microbiology, August 1999, p. 3398-3400, Vol. 65, No. 8
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Ishii, S., Yamamoto, M., Kikuchi, M., Oshima, K., Hattori, M., Otsuka, S., Senoo, K. (2009). Microbial Populations Responsive to Denitrification-Inducing Conditions in Rice Paddy Soil, as Revealed by Comparative 16S rRNA Gene Analysis. Appl. Environ. Microbiol. 75: 7070-7078 [Abstract] [Full Text]  
  • Arthurson, V. (2008). Proper Sanitization of Sewage Sludge: a Critical Issue for a Sustainable Society. Appl. Environ. Microbiol. 74: 5267-5275 [Full Text]  
  • Hamasaki, K., Taniguchi, A., Tada, Y., Long, R. A., Azam, F. (2007). Actively Growing Bacteria in the Inland Sea of Japan, Identified by Combined Bromodeoxyuridine Immunocapture and Denaturing Gradient Gel Electrophoresis. Appl. Environ. Microbiol. 73: 2787-2798 [Abstract] [Full Text]  
  • Schwartz, E. (2007). Characterization of Growing Microorganisms in Soil by Stable Isotope Probing with H218O. Appl. Environ. Microbiol. 73: 2541-2546 [Abstract] [Full Text]  
  • Walters, S. P., Field, K. G. (2006). Persistence and growth of fecal bacteroidales assessed by bromodeoxyuridine immunocapture.. Appl. Environ. Microbiol. 72: 4532-4539 [Abstract] [Full Text]  
  • Liu, X., Bagwell, C. E., Wu, L., Devol, A. H., Zhou, J. (2003). Molecular Diversity of Sulfate-Reducing Bacteria from Two Different Continental Margin Habitats. Appl. Environ. Microbiol. 69: 6073-6081 [Abstract] [Full Text]  
  • Pernthaler, A., Pernthaler, J., Schattenhofer, M., Amann, R. (2002). Identification of DNA-Synthesizing Bacterial Cells in Coastal North Sea Plankton. Appl. Environ. Microbiol. 68: 5728-5736 [Abstract] [Full Text]  
  • Yin, B., Crowley, D., Sparovek, G., De Melo, W. J., Borneman, J. (2000). Bacterial Functional Redundancy along a Soil Reclamation Gradient. Appl. Environ. Microbiol. 66: 4361-4365 [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»