Visualization of Viral Infection Dynamics in a Unicellular Eukaryote and Quantification of Viral Production Using Virus Fluorescence in situ Hybridization

Castillo, Yaiza; Sebastián, Marta; Forn, Irene; Grimsley, Nigel; Yau, Sheree; Moraru, Cristina; Vaqué, Dolors; Ministerio de Economía y Competitividad (España)

Visualization of Viral Infection Dynamics in a Unicellular Eukaryote and Quantification of Viral Production Using Virus Fluorescence in situ Hybridization

11 pages, 7 figures, supplementary material https://www.frontiersin.org/articles/10.3389/fmicb.2020.01559/full#supplementary-material.-- All datasets generated for this study are included in the article/Supplementary Material

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Main Authors:	Castillo, Yaiza, Sebastián, Marta, Forn, Irene, Grimsley, Nigel, Yau, Sheree, Moraru, Cristina, Vaqué, Dolors
Other Authors:	Ministerio de Economía y Competitividad (España)
Format:	artículo biblioteca
Language:	English
Published:	Frontiers Media 2020-07
Subjects:	Virus fluorescence in situ hybridization, Ostreococcus tauri, Ostreococcus tauri virus 5, Virus-host interactions, Culture system, Marine picoeukaryote,
Online Access:	http://hdl.handle.net/10261/217022 http://dx.doi.org/10.13039/501100007757 http://dx.doi.org/10.13039/501100001665 http://dx.doi.org/10.13039/501100003329 http://dx.doi.org/10.13039/501100011033
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spelling	dig-icm-es-10261-2170222021-12-28T15:35:12Z Visualization of Viral Infection Dynamics in a Unicellular Eukaryote and Quantification of Viral Production Using Virus Fluorescence in situ Hybridization Castillo, Yaiza Sebastián, Marta Forn, Irene Grimsley, Nigel Yau, Sheree Moraru, Cristina Vaqué, Dolors Ministerio de Economía y Competitividad (España) Agencia Canaria de Investigación, Innovación y Sociedad de la Información Agence Nationale de la Recherche (France) Agencia Estatal de Investigación (España) Virus fluorescence in situ hybridization Ostreococcus tauri Ostreococcus tauri virus 5 Virus-host interactions Culture system Marine picoeukaryote 11 pages, 7 figures, supplementary material https://www.frontiersin.org/articles/10.3389/fmicb.2020.01559/full#supplementary-material.-- All datasets generated for this study are included in the article/Supplementary Material One of the major challenges in viral ecology is to assess the impact of viruses in controlling the abundance of specific hosts in the environment. To this end, techniques that enable the detection and quantification of virus-host interactions at the single-cell level are essential. With this goal in mind, we implemented virus fluorescence in situ hybridization (VirusFISH) using as a model the marine picoeukaryote Ostreococcus tauri and its virus Ostreococcus tauri virus 5 (OtV5). VirusFISH allowed the visualization and quantification of the proportion of infected cells during an infection cycle in experimental conditions. We were also able to quantify the abundance of free viruses released during cell lysis, discriminating OtV5 from other mid-level fluorescence phages in our non-axenic infected culture that were not easily distinguishable with flow cytometry. Our results showed that although the major lysis of the culture occurred between 24 and 48 h after OtV5 inoculation, some new viruses were already produced between 8 and 24 h. With this work, we demonstrate that VirusFISH is a promising technique to study specific virus-host interactions in non-axenic cultures and establish a framework for its application in complex natural communities YC was supported by Ministerio de Economía, Industria y Competitividad, Gobierno de España (MINECO), grant number: BES-2014-067849 during her PhD. MS was supported by Agencia Canaria de Investigación, Innovación y Sociedad de la Información (ACIISI), grant: Viera y Clavijo 2016. Ministerio de Economía, Industria y Competitividad, Gobierno de España (MINECO), grant numbers: CTM2013‐43767‐P and CTM2016‐75083‐R supported the project and paid for the investigation (reagents, material, etc.). NG was supported by Agence Nationale de la Recherche (ANR), grant number: ANR-17-CE02-0012-03. With the funding support of the ‘Severo Ochoa Centre of Excellence’ accreditation (CEX2019-000928-S), of the Spanish Research Agency (AEI) Peer reviewed 2020-07-27T07:51:16Z 2020-07-27T07:51:16Z 2020-07 artículo http://purl.org/coar/resource_type/c_6501 Frontiers in Microbiology 11: 1559 (2020) CEX2019-000928-S http://hdl.handle.net/10261/217022 10.3389/fmicb.2020.01559 1664-302X http://dx.doi.org/10.13039/501100007757 http://dx.doi.org/10.13039/501100001665 http://dx.doi.org/10.13039/501100003329 http://dx.doi.org/10.13039/501100011033 32765451 en #PLACEHOLDER_PARENT_METADATA_VALUE# #PLACEHOLDER_PARENT_METADATA_VALUE# info:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/CTM2013-43767-P info:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/CTM2016-75083-R Publisher's version https://doi.org/10.3389/fmicb.2020.01559 Sí open Frontiers Media
institution	ICM ES
collection	DSpace
country	España
countrycode	ES
component	Bibliográfico
access	En linea
databasecode	dig-icm-es
tag	biblioteca
region	Europa del Sur
libraryname	Biblioteca del ICM España
language	English
topic	Virus fluorescence in situ hybridization Ostreococcus tauri Ostreococcus tauri virus 5 Virus-host interactions Culture system Marine picoeukaryote Virus fluorescence in situ hybridization Ostreococcus tauri Ostreococcus tauri virus 5 Virus-host interactions Culture system Marine picoeukaryote
spellingShingle	Virus fluorescence in situ hybridization Ostreococcus tauri Ostreococcus tauri virus 5 Virus-host interactions Culture system Marine picoeukaryote Virus fluorescence in situ hybridization Ostreococcus tauri Ostreococcus tauri virus 5 Virus-host interactions Culture system Marine picoeukaryote Castillo, Yaiza Sebastián, Marta Forn, Irene Grimsley, Nigel Yau, Sheree Moraru, Cristina Vaqué, Dolors Visualization of Viral Infection Dynamics in a Unicellular Eukaryote and Quantification of Viral Production Using Virus Fluorescence in situ Hybridization
description	11 pages, 7 figures, supplementary material https://www.frontiersin.org/articles/10.3389/fmicb.2020.01559/full#supplementary-material.-- All datasets generated for this study are included in the article/Supplementary Material
author2	Ministerio de Economía y Competitividad (España)
author_facet	Ministerio de Economía y Competitividad (España) Castillo, Yaiza Sebastián, Marta Forn, Irene Grimsley, Nigel Yau, Sheree Moraru, Cristina Vaqué, Dolors
format	artículo
topic_facet	Virus fluorescence in situ hybridization Ostreococcus tauri Ostreococcus tauri virus 5 Virus-host interactions Culture system Marine picoeukaryote
author	Castillo, Yaiza Sebastián, Marta Forn, Irene Grimsley, Nigel Yau, Sheree Moraru, Cristina Vaqué, Dolors
author_sort	Castillo, Yaiza
title	Visualization of Viral Infection Dynamics in a Unicellular Eukaryote and Quantification of Viral Production Using Virus Fluorescence in situ Hybridization
title_short	Visualization of Viral Infection Dynamics in a Unicellular Eukaryote and Quantification of Viral Production Using Virus Fluorescence in situ Hybridization
title_full	Visualization of Viral Infection Dynamics in a Unicellular Eukaryote and Quantification of Viral Production Using Virus Fluorescence in situ Hybridization
title_fullStr	Visualization of Viral Infection Dynamics in a Unicellular Eukaryote and Quantification of Viral Production Using Virus Fluorescence in situ Hybridization
title_full_unstemmed	Visualization of Viral Infection Dynamics in a Unicellular Eukaryote and Quantification of Viral Production Using Virus Fluorescence in situ Hybridization
title_sort	visualization of viral infection dynamics in a unicellular eukaryote and quantification of viral production using virus fluorescence in situ hybridization
publisher	Frontiers Media
publishDate	2020-07
url	http://hdl.handle.net/10261/217022 http://dx.doi.org/10.13039/501100007757 http://dx.doi.org/10.13039/501100001665 http://dx.doi.org/10.13039/501100003329 http://dx.doi.org/10.13039/501100011033
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Visualization of Viral Infection Dynamics in a Unicellular Eukaryote and Quantification of Viral Production Using Virus Fluorescence in situ Hybridization

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