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Applied and Environmental Microbiology, November 2004, p. 6542-6550, Vol. 70, No. 11
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.11.6542-6550.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Representative Freshwater Bacterioplankton Isolated from Crater Lake, Oregon

Kathleen A. Page,1* Stephanie A. Connon,2,{dagger} and Stephen J. Giovannoni2

Department of Biology, Southern Oregon University, Ashland,1 Department of Microbiology, Oregon State University, Corvallis, Oregon2

Received 27 April 2004/ Accepted 4 July 2004

High-throughput culturing (HTC) methods that rely on dilution to extinction in very-low-nutrient media were used to obtain bacterial isolates from Crater Lake, Oregon. 16S rRNA sequence determination and phylogenetic reconstruction were used to determine the potential ecological significance of isolated bacteria, both in Crater Lake and globally. Fifty-five Crater Lake isolates yielded 16 different 16S rRNA gene sequences. Thirty of 55 (55%) Crater Lake isolates had 16S rRNA gene sequences with 97% or greater similarity to sequences recovered previously from Crater Lake 16S rRNA gene clone libraries. Furthermore, 36 of 55 (65%) Crater Lake isolates were found to be members of widely distributed freshwater groups. These results confirm that HTC is a significant improvement over traditional isolation techniques that tend to enrich for microorganisms that do not predominate in their environment and rarely correlate with 16S rRNA gene clone library sequences. Although all isolates were obtained under dark, heterotrophic growth conditions, 2 of the 16 different groups showed evidence of photosynthetic capability as assessed by the presence of puf operon sequences, suggesting that photoheterotrophy may be a significant process in this oligotrophic, freshwater habitat.

* Corresponding author. Mailing address: Dept. of Biology, Southern Oregon University, 1250 Siskiyou Blvd., Ashland, OR 97520. Phone: (541) 552-6788. Fax: (541) 552-6415. E-mail: page{at}sou.edu.

{dagger} Present address: Department of Biology, Idaho State University, Pocatello, ID 83209.

Applied and Environmental Microbiology, November 2004, p. 6542-6550, Vol. 70, No. 11
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.11.6542-6550.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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