Background. Viruses strongly influence microbial population dynamics and ecosystem functions However, our ability to quantitatively evaluate those viral impads is limited to the few cultivated viruses and double-stranded DNA (dsDNA) viral genomes captured in quantitative viral metagenornes (vromes). This leaves the ecology of nondsDNA viruses nearly unlmovvn, including single-stranded DNA (ssDNA) viruses that have been frequently observed in viromes, but not quantified due to amplification biases in sequencing library preparations (Multiple Displacement Amplification, Linker Amplification or Tagmentation). Methods. Here we designed mock viral communities including both ssDNA and dsDNA viruses to evaluate the capability of a sequencing library preparation approach including an Adaptase step prior to Linker Amplification for quantitative amplification of both dsDNA and ssDNA templates. We then surveyed aquatic samples to provide first estimates of the abundance of ssDNA viruses. Results. Mock community experiments confirmed the biased nature of existing library preparation methods for ssDNA templates (either largely enriched or selected against) and showed that the protocol using Adaptase plus Linker Amplification yielded viromes that were 1.8-fold quantitative for ssDNA and dsDNA viruses. Application of this protocol to community virus DNA from three freshwater and three marine samples revealed that ssDNA viruses as a whole represent only a minor fraction (<5%) of DNA virus communities, though individual ssDNA genomes, both eukaryoteinfecting Circular Rep-Encoding Single-Stranded DNA (CRESS-DNA) viruses and bacteriophages from the Microviridae family, can be among the most abundant viral genomes in a sample. Discussion. Together these findings provide empirical data for a new virome library preparation protocol, and a first estimate of ssDNA virus abundance in aquatic systems.
| Identifer | oai:union.ndltd.org:arizona.edu/oai:arizona.openrepository.com:10150/622482 |
| Date | 08 December 2016 |
| Creators | Roux, Simon, Solonenko, Natalie E., Dang, Vinh T., Poulos, Bonnie T., Schwenck, Sarah M., Goldsmith, Dawn B., Coleman, Maureen L., Breitbart, Mya, Sullivan, Matthew B. |
| Contributors | Univ Arizona, Dept Ecol & Evolutionary Biol, Department of Microbiology, The Ohio State University, Columbus, OH, United States, Department of Microbiology, The Ohio State University, Columbus, OH, United States, Department of Microbiology, Ha Long University, Uong Bi, Quang Ninh, Vietnam, Department of Ecology and Evolutionary Biology, University of Arizona, Tucson, AZ, United States, Department of Ecology and Evolutionary Biology, University of Arizona, Tucson, AZ, United States, College of Marine Science, University of South Florida, St. Petersburg, FL, United States, Department of the Geophysical Sciences, University of Chicago, Chicago, IL, United States, College of Marine Science, University of South Florida, St. Petersburg, FL, United States, Department of Microbiology, The Ohio State University, Columbus, OH, United States |
| Publisher | PEERJ INC |
| Source Sets | University of Arizona |
| Language | English |
| Detected Language | English |
| Type | Article |
| Rights | © 2016 Roux et al. Distributed under Creative Commons CC-BY 4.0 |
| Relation | https://peerj.com/articles/2777 |
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