Isolation and Genetic Analysis of Haloalkaliphilic Bacteriophages in a North American Soda Lake
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Mono Lake is a meromictic, hypersaline, soda lake that harbors a diverse and abundant microbial community. A previous report documented the high viral abundance in Mono Lake, and pulsed-field gel electrophoresis analysis of viral DNA from lake water samples showed a diverse population based on a broad range of viral genome sizes. To better understand the ecology of bacteriophages and their hosts in this unique environment, water samples were collected between February 2001 and July 2004 for isolation of bacteriophages by using four indigenous bacterial hosts. Plaque assay results showed a differential seasonal expression of cultured bacteriophages. To reveal the diversity of uncultured bacteriophages, viral DNA from lake water samples was used to construct clone libraries. Sequence analysis of viral clones revealed homology to viral as well as bacterial proteins. Furthermore, dot blot DNA hybridization analyses showed that the uncultured viruses are more prevalent during most seasons, whereas the viral isolates (Aφ and φ2) were less prevalent, confirming the belief that uncultured viruses represent the dominant members of the community, whereas cultured isolates represent the minority species.
KeywordsBacterial Artificial Chromosome Bacterial Artificial Chromosome Clone Soda Lake Bacterial Host Mono Lake
We thank Dr. Grieg Steward and Dr. Robert Jellison for their technical expertise and their collaboration in field sampling. We also thank Sandra Roll, Kimberly Rose, and other members of the SNARL team for collecting water samples during this study. We thank Sam Choi for his help with statistical analysis and anonymous reviewers for their suggestions for improving this manuscript. This study was supported by NSF awards DEB-01-30528, DEB-01-29174, and DEB-01-29160 to S.C.J., G.F.S., and R.J., respectively.
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