Abstract
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.
Highlights
Marine viruses have been studied during the last 3 decades mostly using microscopy (Noble and Fuhrman, 1998) and flow cytometry (FCM; Marie et al, 1999) for the enumeration and estimation of viral production
Information about the viral taxonomic and genomic diversity, about their biogeography and, to a certain extent, about their potential hosts (e.g., Chow et al, 2015; Labonté et al, 2015; Castillo et al, 2019). They do not allow the visualization of specific virus-host interactions and the monitoring of infection dynamics, which are crucial to better understand the role of viruses in shaping microbial communities and biogeochemical cycles
A non-axenic culture of O. tauri was infected with the virus Ostreococcus tauri virus 5 (OtV5), at a multiplicity of infection (MOI) of 0.01 and an uninfected culture was grown in parallel, as a control (Figure 2A)
Summary
Marine viruses have been studied during the last 3 decades mostly using microscopy (Noble and Fuhrman, 1998) and flow cytometry (FCM; Marie et al, 1999) for the enumeration and estimation of viral production. O. tauri Infection Dynamics Through VirusFISH information about the viral taxonomic and genomic diversity, about their biogeography and, to a certain extent, about their potential hosts (e.g., Chow et al, 2015; Labonté et al, 2015; Castillo et al, 2019). They do not allow the visualization of specific virus-host interactions and the monitoring of infection dynamics, which are crucial to better understand the role of viruses in shaping microbial communities and biogeochemical cycles. They managed to directly detect the phagehost association by targeting genes from both components without culturing but with no visual representation of the infection
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