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      Quantitative Viral Community DNA Analysis Reveals the Dominance of Single-Stranded DNA Viruses in Offshore Upper Bathyal Sediment from Tohoku, Japan

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          Abstract

          Previous studies on marine environmental virology have primarily focused on double-stranded DNA (dsDNA) viruses; however, it has recently been suggested that single-stranded DNA (ssDNA) viruses are more abundant in marine ecosystems. In this study, we performed a quantitative viral community DNA analysis to estimate the relative abundance and composition of both ssDNA and dsDNA viruses in offshore upper bathyal sediment from Tohoku, Japan (water depth = 500 m). The estimated dsDNA viral abundance ranged from 3 × 10 6 to 5 × 10 6 genome copies per cm 3 sediment, showing values similar to the range of fluorescence-based direct virus counts. In contrast, the estimated ssDNA viral abundance ranged from 1 × 10 8 to 3 × 10 9 genome copies per cm 3 sediment, thus providing an estimation that the ssDNA viral populations represent 96.3–99.8% of the benthic total DNA viral assemblages. In the ssDNA viral metagenome, most of the identified viral sequences were associated with ssDNA viral families such as Circoviridae and Microviridae. The principle components analysis of the ssDNA viral sequence components from the sedimentary ssDNA viral metagenomic libraries found that the different depth viral communities at the study site all exhibited similar profiles compared with deep-sea sediment ones at other reference sites. Our results suggested that deep-sea benthic ssDNA viruses have been significantly underestimated by conventional direct virus counts and that their contributions to deep-sea benthic microbial mortality and geochemical cycles should be further addressed by such a new quantitative approach.

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          Virioplankton: Viruses in Aquatic Ecosystems

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            Functional metagenomic profiling of nine biomes.

            Microbial activities shape the biogeochemistry of the planet and macroorganism health. Determining the metabolic processes performed by microbes is important both for understanding and for manipulating ecosystems (for example, disruption of key processes that lead to disease, conservation of environmental services, and so on). Describing microbial function is hampered by the inability to culture most microbes and by high levels of genomic plasticity. Metagenomic approaches analyse microbial communities to determine the metabolic processes that are important for growth and survival in any given environment. Here we conduct a metagenomic comparison of almost 15 million sequences from 45 distinct microbiomes and, for the first time, 42 distinct viromes and show that there are strongly discriminatory metabolic profiles across environments. Most of the functional diversity was maintained in all of the communities, but the relative occurrence of metabolisms varied, and the differences between metagenomes predicted the biogeochemical conditions of each environment. The magnitude of the microbial metabolic capabilities encoded by the viromes was extensive, suggesting that they serve as a repository for storing and sharing genes among their microbial hosts and influence global evolutionary and metabolic processes.
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              Rapid amplification of plasmid and phage DNA using Phi 29 DNA polymerase and multiply-primed rolling circle amplification.

              We describe a simple method of using rolling circle amplification to amplify vector DNA such as M13 or plasmid DNA from single colonies or plaques. Using random primers and phi29 DNA polymerase, circular DNA templates can be amplified 10,000-fold in a few hours. This procedure removes the need for lengthy growth periods and traditional DNA isolation methods. Reaction products can be used directly for DNA sequencing after phosphatase treatment to inactivate unincorporated nucleotides. Amplified products can also be used for in vitro cloning, library construction, and other molecular biology applications.
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                Author and article information

                Contributors
                Journal
                Front Microbiol
                Front Microbiol
                Front. Microbiol.
                Frontiers in Microbiology
                Frontiers Media S.A.
                1664-302X
                06 February 2018
                2018
                : 9
                : 75
                Affiliations
                [1] 1Department of Subsurface Geobiological Analysis and Research, Japan Agency for Marine-Earth Science and Technology , Yokosuka, Japan
                [2] 2Project Team for Analyses of Changes in East Japan Marine Ecosystems, Japan Agency for Marine-Earth Science and Technology , Yokosuka, Japan
                [3] 3Earth-Life Science Institute, Tokyo Institute of Technology , Tokyo, Japan
                [4] 4Research and Development Center for Marine Biosciences, Japan Agency for Marine-Earth Science and Technology , Yokosuka, Japan
                [5] 5Department of Biogeochemistry, Japan Agency for Marine-Earth Science and Technology , Yokosuka, Japan
                Author notes

                Edited by: Feng Chen, University of Maryland Center for Environmental Science, United States

                Reviewed by: Jessica Labonté, Texas A&M University Galveston Campus, United States; Kui Wang, Institute of Marine and Environmental Technology, University of Maryland Center for Environmental Science, United States

                *Correspondence: Mitsuhiro Yoshida, mityoshi@ 123456jamstec.go.jp

                This article was submitted to Aquatic Microbiology, a section of the journal Frontiers in Microbiology

                Article
                10.3389/fmicb.2018.00075
                5807898
                29467725
                1a61e6e4-e2a9-4fb6-ba84-28cb9476df1d
                Copyright © 2018 Yoshida, Mochizuki, Urayama, Yoshida-Takashima, Nishi, Hirai, Nomaki, Takaki, Nunoura and Takai.

                This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

                History
                : 04 October 2017
                : 12 January 2018
                Page count
                Figures: 4, Tables: 2, Equations: 0, References: 52, Pages: 10, Words: 0
                Funding
                Funded by: Japan Society for the Promotion of Science 10.13039/501100001691
                Award ID: 26660154
                Funded by: Ministry of Education, Culture, Sports, Science and Technology 10.13039/501100001700
                Award ID: 16H06429
                Award ID: 16K21723
                Award ID: 17H05830
                Award ID: 16H06437
                Categories
                Microbiology
                Original Research

                Microbiology & Virology
                deep-sea sediment,single-stranded dna viruses,genome quantification,marine microbiology,metagenomics

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