This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Santegoeds, C. M. Articles by Ward, D. M. Search for Related Content PubMed PubMed Citation Articles by Santegoeds, C. M. Articles by Ward, D. M. Agricola Articles by Santegoeds, C. M. Articles by Ward, D. M.

 Previous Article  |  Next Article 

Appl. Environ. Microbiol., Nov 1996, 3922-3928, Vol 62, No. 11
Copyright © 1996, American Society for Microbiology

Denaturing gradient gel electrophoresis used to monitor the enrichment culture of aerobic chemoorganotrophic bacteria from a hot spring cyanobacterial mat

CM Santegoeds, SC Nold and DM Ward
Department of Microbiology, Montana State University, Bozeman 59717, USA.

Previous studies investigating microbial diversity in the Octopus Spring cyanobacterial mat community (Yellowstone National Park) have shown a discrepancy between bacterial populations observed by molecular retrieval and cultivation techniques. To investigate how selective enrichment culture techniques affect species composition, we used denaturing gradient gel electrophoresis (DGGE) separation of PCR- amplified 16S rRNA gene fragments to monitor the populations contained within enrichment cultures of aerobic chemoorganotrophic bacteria from the ca. 50 degrees C region of the mat community. By varying the degree of dilution of the inoculum, medium composition, and enrichment conditions and duration and by analyzing the cultures by DGGE, we detected 14 unique 16S rRNA sequence types. These corresponded to alpha- , beta-, gamma-, and delta-proteobacteria, Thermus relatives, and gram- positive bacteria with high G + C ratio and, at the highest inoculum dilutions, Chloroflexus aurantiacus relatives, which were estimated to still be approximately 300 times less abundant than cells of the mat primary producer, Synechococcus spp. Only three of these populations were previously cultivated on solidified medium after similar enrichment. Only two of these population have 16S rRNA sequences which were previously cloned directly from the mat. These results reveal a diversity of bacterial populations in enrichment culture which were not detected by either molecular retrieval or strain purification techniques.

This article has been cited by other articles:

• Sousa, D. Z., Pereira, M. A., Stams, A. J. M., Alves, M. M., Smidt, H. (2007). Microbial Communities Involved in Anaerobic Degradation of Unsaturated or Saturated Long-Chain Fatty Acids. Appl. Environ. Microbiol. 73: 1054-1064 [Abstract] [Full Text]  
• Li, Y., Ku, C.Y.S., Xu, J., Saxena, D., Caufield, P.W. (2005). Survey of Oral Microbial Diversity using PCR-based Denaturing Gradient Gel Electrophoresis. J. Dent. Res. 84: 559-564 [Abstract] [Full Text]  
• Hobel, C. F. V., Marteinsson, V. T., Hreggvidsson, G. O., Kristjansson, J. K. (2005). Investigation of the Microbial Ecology of Intertidal Hot Springs by Using Diversity Analysis of 16S rRNA and Chitinase Genes. Appl. Environ. Microbiol. 71: 2771-2776 [Abstract] [Full Text]  
• Morris, C. E., Bardin, M., Berge, O., Frey-Klett, P., Fromin, N., Girardin, H., Guinebretiere, M.-H., Lebaron, P., Thiery, J. M., Troussellier, M. (2002). Microbial Biodiversity: Approaches to Experimental Design and Hypothesis Testing in Primary Scientific Literature from 1975 to 1999. Microbiol. Mol. Biol. Rev. 66: 592-616 [Abstract] [Full Text]  
• Heidelberg, J. F., Heidelberg, K. B., Colwell, R. R. (2002). Seasonality of Chesapeake Bay Bacterioplankton Species. Appl. Environ. Microbiol. 68: 5488-5497 [Abstract] [Full Text]  
• Nubel, U., Bateson, M. M., Vandieken, V., Wieland, A., Kuhl, M., Ward, D. M. (2002). Microscopic Examination of Distribution and Phenotypic Properties of Phylogenetically Diverse Chloroflexaceae-Related Bacteria in Hot Spring Microbial Mats. Appl. Environ. Microbiol. 68: 4593-4603 [Abstract] [Full Text]  
• Ogier, J.-C., Son, O., Gruss, A., Tailliez, P., Delacroix-Buchet, A. (2002). Identification of the Bacterial Microflora in Dairy Products by Temporal Temperature Gradient Gel Electrophoresis. Appl. Environ. Microbiol. 68: 3691-3701 [Abstract] [Full Text]  
• Pinar, G., Ramos, C., Rolleke, S., Schabereiter-Gurtner, C., Vybiral, D., Lubitz, W., Denner, E. B. M. (2001). Detection of Indigenous Halobacillus Populations in Damaged Ancient Wall Paintings and Building Materials: Molecular Monitoring and Cultivation. Appl. Environ. Microbiol. 67: 4891-4895 [Abstract] [Full Text]  
• Burton, S. A. Q., Prosser, J. I. (2001). Autotrophic Ammonia Oxidation at Low pH through Urea Hydrolysis. Appl. Environ. Microbiol. 67: 2952-2957 [Abstract] [Full Text]  
• El-Fantroussi, S. (2000). Enrichment and Molecular Characterization of a Bacterial Culture That Degrades Methoxy-Methyl Urea Herbicides and Their Aniline Derivatives. Appl. Environ. Microbiol. 66: 5110-5115 [Abstract] [Full Text]  
• Friedrich, M., Grosser, R. J., Kern, E. A., Inskeep, W. P., Ward, D. M. (2000). Effect of Model Sorptive Phases on Phenanthrene Biodegradation: Molecular Analysis of Enrichments and Isolates Suggests Selection Based on Bioavailability. Appl. Environ. Microbiol. 66: 2703-2710 [Abstract] [Full Text]  
• Casamayor, E. O., Schäfer, H., Bañeras, L., Pedrós-Alió, C., Muyzer, G. (2000). Identification of and Spatio-Temporal Differences between Microbial Assemblages from Two Neighboring Sulfurous Lakes: Comparison by Microscopy and Denaturing Gradient Gel Electrophoresis. Appl. Environ. Microbiol. 66: 499-508 [Abstract] [Full Text]  
• Heuer, H., Hartung, K., Wieland, G., Kramer, I., Smalla, K. (1999). Polynucleotide Probes That Target a Hypervariable Region of 16S rRNA Genes To Identify Bacterial Isolates Corresponding to Bands of Community Fingerprints. Appl. Environ. Microbiol. 65: 1045-1049 [Abstract] [Full Text]  
• Eichner, C. A., Erb, R. W., Timmis, K. N., Wagner-Döbler, I. (1999). Thermal Gradient Gel Electrophoresis Analysis of Bioprotection from Pollutant Shocks in the Activated Sludge Microbial Community. Appl. Environ. Microbiol. 65: 102-109 [Abstract] [Full Text]  
• Brinkhoff, T., Santegoeds, C. M., Sahm, K., Kuever, J., Muyzer, G. (1998). A Polyphasic Approach To Study the Diversity and Vertical Distribution of Sulfur-Oxidizing Thiomicrospira Species in Coastal Sediments of the German Wadden Sea. Appl. Environ. Microbiol. 64: 4650-4657 [Abstract] [Full Text]  
• Schwieger, F., Tebbe, C. C. (1998). A New Approach To Utilize PCR-Single-Strand-Conformation Polymorphism for 16S rRNA Gene-Based Microbial Community Analysis. Appl. Environ. Microbiol. 64: 4870-4876 [Abstract] [Full Text]  
• Jackson, C. R., Roden, E. E., Churchill, P. F. (1998). Changes in Bacterial Species Composition in Enrichment Cultures with Various Dilutions of Inoculum as Monitored by Denaturing Gradient Gel Electrophoresis. Appl. Environ. Microbiol. 64: 5046-5048 [Abstract] [Full Text]  
• Ward, D. M., Ferris, M. J., Nold, S. C., Bateson, M. M. (1998). A Natural View of Microbial Biodiversity within Hot Spring Cyanobacterial Mat Communities. Microbiol. Mol. Biol. Rev. 62: 1353-1370 [Abstract] [Full Text]  
• Stephen, J. R., Kowalchuk, G. A., Bruns, M.-A. V., McCaig, A. E., Phillips, C. J., Embley, T. M., Prosser, J. I. (1998). Analysis of beta -Subgroup Proteobacterial Ammonia Oxidizer Populations in Soil by Denaturing Gradient Gel Electrophoresis Analysis and Hierarchical Phylogenetic Probing. Appl. Environ. Microbiol. 64: 2958-2965 [Abstract] [Full Text]  
• Großkopf, R., Janssen, P. H., Liesack, W. (1998). Diversity and Structure of the Methanogenic Community in Anoxic Rice Paddy Soil Microcosms as Examined by Cultivation and Direct 16S rRNA Gene Sequence Retrieval. Appl. Environ. Microbiol. 64: 960-969 [Abstract] [Full Text]