Towards optimized viral metagenomes for double-stranded and single-stranded DNA viruses from challenging soils
Soils impact global carbon cycling and their resident microbes are critical to their biogeochemical processing and ecosystem outputs. Based on studies in marine systems, viruses infecting soil microbes likely modulate host activities via mortality, horizontal gene transfer, and metabolic control. Ho...
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ftdoajarticles:oai:doaj.org/article:36ba41b4e0d44090970f5e501b79f697 2024-01-07T09:45:59+01:00 Towards optimized viral metagenomes for double-stranded and single-stranded DNA viruses from challenging soils Gareth Trubl Simon Roux Natalie Solonenko Yueh-Fen Li Benjamin Bolduc Josué Rodríguez-Ramos Emiley A. Eloe-Fadrosh Virginia I. Rich Matthew B. Sullivan 2019-07-01T00:00:00Z https://doi.org/10.7717/peerj.7265 https://doaj.org/article/36ba41b4e0d44090970f5e501b79f697 EN eng PeerJ Inc. https://peerj.com/articles/7265.pdf https://peerj.com/articles/7265/ https://doaj.org/toc/2167-8359 doi:10.7717/peerj.7265 2167-8359 https://doaj.org/article/36ba41b4e0d44090970f5e501b79f697 PeerJ, Vol 7, p e7265 (2019) Soil viruses Viromes DNA extraction Organics Microbiology ssDNA viruses Medicine R Biology (General) QH301-705.5 article 2019 ftdoajarticles https://doi.org/10.7717/peerj.7265 2023-12-10T01:51:02Z Soils impact global carbon cycling and their resident microbes are critical to their biogeochemical processing and ecosystem outputs. Based on studies in marine systems, viruses infecting soil microbes likely modulate host activities via mortality, horizontal gene transfer, and metabolic control. However, their roles remain largely unexplored due to technical challenges with separating, isolating, and extracting DNA from viruses in soils. Some of these challenges have been overcome by using whole genome amplification methods and while these have allowed insights into the identities of soil viruses and their genomes, their inherit biases have prevented meaningful ecological interpretations. Here we experimentally optimized steps for generating quantitatively-amplified viral metagenomes to better capture both ssDNA and dsDNA viruses across three distinct soil habitats along a permafrost thaw gradient. First, we assessed differing DNA extraction methods (PowerSoil, Wizard mini columns, and cetyl trimethylammonium bromide) for quantity and quality of viral DNA. This established PowerSoil as best for yield and quality of DNA from our samples, though ∼1/3 of the viral populations captured by each extraction kit were unique, suggesting appreciable differential biases among DNA extraction kits. Second, we evaluated the impact of purifying viral particles after resuspension (by cesium chloride gradients; CsCl) and of viral lysis method (heat vs bead-beating) on the resultant viromes. DNA yields after CsCl particle-purification were largely non-detectable, while unpurified samples yielded 1–2-fold more DNA after lysis by heat than by bead-beating. Virome quality was assessed by the number and size of metagenome-assembled viral contigs, which showed no increase after CsCl-purification, but did from heat lysis relative to bead-beating. We also evaluated sample preparation protocols for ssDNA virus recovery. In both CsCl-purified and non-purified samples, ssDNA viruses were successfully recovered by using the Accel-NGS 1S ... Article in Journal/Newspaper permafrost Directory of Open Access Journals: DOAJ Articles PeerJ 7 e7265 |
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ftdoajarticles |
language |
English |
topic |
Soil viruses Viromes DNA extraction Organics Microbiology ssDNA viruses Medicine R Biology (General) QH301-705.5 |
spellingShingle |
Soil viruses Viromes DNA extraction Organics Microbiology ssDNA viruses Medicine R Biology (General) QH301-705.5 Gareth Trubl Simon Roux Natalie Solonenko Yueh-Fen Li Benjamin Bolduc Josué Rodríguez-Ramos Emiley A. Eloe-Fadrosh Virginia I. Rich Matthew B. Sullivan Towards optimized viral metagenomes for double-stranded and single-stranded DNA viruses from challenging soils |
topic_facet |
Soil viruses Viromes DNA extraction Organics Microbiology ssDNA viruses Medicine R Biology (General) QH301-705.5 |
description |
Soils impact global carbon cycling and their resident microbes are critical to their biogeochemical processing and ecosystem outputs. Based on studies in marine systems, viruses infecting soil microbes likely modulate host activities via mortality, horizontal gene transfer, and metabolic control. However, their roles remain largely unexplored due to technical challenges with separating, isolating, and extracting DNA from viruses in soils. Some of these challenges have been overcome by using whole genome amplification methods and while these have allowed insights into the identities of soil viruses and their genomes, their inherit biases have prevented meaningful ecological interpretations. Here we experimentally optimized steps for generating quantitatively-amplified viral metagenomes to better capture both ssDNA and dsDNA viruses across three distinct soil habitats along a permafrost thaw gradient. First, we assessed differing DNA extraction methods (PowerSoil, Wizard mini columns, and cetyl trimethylammonium bromide) for quantity and quality of viral DNA. This established PowerSoil as best for yield and quality of DNA from our samples, though ∼1/3 of the viral populations captured by each extraction kit were unique, suggesting appreciable differential biases among DNA extraction kits. Second, we evaluated the impact of purifying viral particles after resuspension (by cesium chloride gradients; CsCl) and of viral lysis method (heat vs bead-beating) on the resultant viromes. DNA yields after CsCl particle-purification were largely non-detectable, while unpurified samples yielded 1–2-fold more DNA after lysis by heat than by bead-beating. Virome quality was assessed by the number and size of metagenome-assembled viral contigs, which showed no increase after CsCl-purification, but did from heat lysis relative to bead-beating. We also evaluated sample preparation protocols for ssDNA virus recovery. In both CsCl-purified and non-purified samples, ssDNA viruses were successfully recovered by using the Accel-NGS 1S ... |
format |
Article in Journal/Newspaper |
author |
Gareth Trubl Simon Roux Natalie Solonenko Yueh-Fen Li Benjamin Bolduc Josué Rodríguez-Ramos Emiley A. Eloe-Fadrosh Virginia I. Rich Matthew B. Sullivan |
author_facet |
Gareth Trubl Simon Roux Natalie Solonenko Yueh-Fen Li Benjamin Bolduc Josué Rodríguez-Ramos Emiley A. Eloe-Fadrosh Virginia I. Rich Matthew B. Sullivan |
author_sort |
Gareth Trubl |
title |
Towards optimized viral metagenomes for double-stranded and single-stranded DNA viruses from challenging soils |
title_short |
Towards optimized viral metagenomes for double-stranded and single-stranded DNA viruses from challenging soils |
title_full |
Towards optimized viral metagenomes for double-stranded and single-stranded DNA viruses from challenging soils |
title_fullStr |
Towards optimized viral metagenomes for double-stranded and single-stranded DNA viruses from challenging soils |
title_full_unstemmed |
Towards optimized viral metagenomes for double-stranded and single-stranded DNA viruses from challenging soils |
title_sort |
towards optimized viral metagenomes for double-stranded and single-stranded dna viruses from challenging soils |
publisher |
PeerJ Inc. |
publishDate |
2019 |
url |
https://doi.org/10.7717/peerj.7265 https://doaj.org/article/36ba41b4e0d44090970f5e501b79f697 |
genre |
permafrost |
genre_facet |
permafrost |
op_source |
PeerJ, Vol 7, p e7265 (2019) |
op_relation |
https://peerj.com/articles/7265.pdf https://peerj.com/articles/7265/ https://doaj.org/toc/2167-8359 doi:10.7717/peerj.7265 2167-8359 https://doaj.org/article/36ba41b4e0d44090970f5e501b79f697 |
op_doi |
https://doi.org/10.7717/peerj.7265 |
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PeerJ |
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7 |
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e7265 |
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1787427655091486720 |