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Cultivation-Based Assessment of Lysogeny Among Soil Bacteria

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Abstract

Lysogeny has long been proposed as an important long-term maintenance strategy for autochthonous soil bacteriophages (phages). Whole genome sequence data indicate that prophage-derived sequences pervade prokaryotic genomes, but the connection between inferred prophage sequence and an active temperate phage is tenuous. Thus, definitive evidence of phage production from lysogenic prokaryotes will be critical in determining the presence and extent of temperate phage diversity existing as prophage within bacterial genomes and within environmental contexts such as soils. This study optimized methods for systematic and definitive determination of lysogeny within a collection of autochthonous soil bacteria. Twenty bacterial isolates from a range of Delaware soil environments (five from each soil) were treated with the inducing agents mitomycin C (MC) or UV light. Six isolates (30%) carried inducible temperate phages as evidenced by an increase in virus direct counts. The magnitude of induction response was highly dependent upon specific induction conditions, and corresponding burst sizes ranged from 1 to 176. Treatment with MC for 30 min yielded the largest induction responses for three of the six lysogens. Morphological analysis revealed that four of the lysogens produced lambda-like Siphoviridae particles, whereas two produced Myoviridae particles. Additionally, pulsed-field gel electrophoresis data indicated that two of the six lysogens were polylysogens, producing more than one distinct type of phage particle. These results suggest that lysogeny is relatively common among soil bacteria.

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Acknowledgment

We wish to thank Kirk Czymmek and Debbie Powell of the Delaware Biotechnology Institute Bio-Imaging Center for technical assistance with electron microscopy, Yingbo Wang for her help with induction assays, and Danielle Winget for reading the manuscript. This work was supported by EPA STAR Graduate Fellowship U916129 to K. E. Williamson and the National Research Initiative of the USDA Cooperative State Research, Education, and Extension Service grant 2005-35107-15214 to K. E. Wommack.

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Authors and Affiliations

  1. Environmental Genomics Group, J. Craig Venter Institute, La Jolla, CA, 92037, USA

    Kurt E. Williamson

  2. Department of Plant and Soil Sciences, University of Delaware, Newark, DE, 19716, USA

    Kurt E. Williamson & K. Eric Wommack

  3. Department of Chemistry and Biochemistry, University of Delaware, Newark, DE, 19716, USA

    Jennifer B. Schnitker

  4. Department of Biosystems Engineering and Soil Science, University of Tennessee, Knoxville, TN, 37996-4531, USA

    Mark Radosevich

  5. Center for Biomarker Analysis, University of Tennessee, Knoxville, TN, 37996, USA

    Mark Radosevich

  6. Department of Biological Sciences, University of Delaware, Newark, DE, 19716, USA

    David W. Smith

  7. Delaware Biotechnology Institute, 15 Innovation Way Lab 148, Newark, DE, 19711, USA

    K. Eric Wommack

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  1. Kurt E. Williamson
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Correspondence to K. Eric Wommack.

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Physical properties of soils (PDF 37.2 KB)

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Williamson, K.E., Schnitker, J.B., Radosevich, M. et al. Cultivation-Based Assessment of Lysogeny Among Soil Bacteria. Microb Ecol 56, 437–447 (2008). https://doi.org/10.1007/s00248-008-9362-2

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