Castillo Yaiza M, Sebastián Marta, Forn Irene, Grimsley Nigel, Yau Sheree, Moraru Cristina, Vaqué Dolors
Department of Marine Biology and Oceanography, Institute of Marine Sciences (CSIC), Barcelona, Spain.
Institute of Oceanography and Global Change (IOCAG), University of Las Palmas de Gran Canaria (ULPGC), Las Palmas de Gran Canaria, Spain.
Front Microbiol. 2020 Jul 17;11:1559. doi: 10.3389/fmicb.2020.01559. eCollection 2020.
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 hybridization (VirusFISH) using as a model the marine picoeukaryote and its virus 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.
病毒生态学的主要挑战之一是评估病毒对控制环境中特定宿主数量的影响。为此，能够在单细胞水平检测和量化病毒-宿主相互作用的技术至关重要。出于这一目标，我们以海洋微微型真核生物及其病毒OtV5为模型实施了病毒荧光杂交（VirusFISH）。VirusFISH使我们能够在实验条件下观察和量化感染周期中受感染细胞的比例。我们还能够量化细胞裂解过程中释放的游离病毒的数量，在我们的非无菌感染培养物中，将OtV5与其他中等荧光强度的噬菌体区分开来，而这些噬菌体用流式细胞术不易区分。我们的结果表明，虽然培养物的主要裂解发生在接种OtV5后的24至48小时之间，但在8至24小时之间已经产生了一些新病毒。通过这项工作，我们证明VirusFISH是研究非无菌培养物中特定病毒-宿主相互作用的一种有前景的技术，并为其在复杂自然群落中的应用建立了一个框架。
