Metaviromes Reveal the Dynamics of <i>Pseudomonas</i> Host-Specific Phages Cultured and Uncultured by Plaque Assay
2021
Katrine Wacenius Skov Alanin | Laura Milena Forero Junco | Jacob Bruun Jørgensen | Tue Kjærgaard Nielsen | Morten Arendt Rasmussen | Witold Kot | Lars Hestbjerg Hansen
Isolating single phages using plaque assays is a laborious and time-consuming process. Whether single isolated phages are the most lyse-effective, the most abundant in viromes, or those with the highest ability to make plaques in solid media is not well known. With the increasing accessibility of high-throughput sequencing, metaviromics is often used to describe viruses in environmental samples. By extracting and sequencing metaviromes from organic waste with and without exposure to a host-of-interest, we show a host-related phage community’s shift, as well as identify the most enriched phages. Moreover, we isolated plaque-forming single phages using the same virome–host matrix to observe how enrichments in liquid media correspond to the metaviromic data. In this study, we observed a significant shift (<i>p</i> = 0.015) of the 47 identified putative <i>Pseudomonas</i> phages with a minimum twofold change above zero in read abundance when adding a <i>Pseudomonas</i> <i>syringae</i> DC3000 host. Surprisingly, it appears that only two out of five plaque-forming phages from the same organic waste sample, targeting the <i>Pseudomonas</i> strain, were highly abundant in the metavirome, while the other three were almost absent despite host exposure. Lastly, our sequencing results highlight how long reads from Oxford Nanopore elevates the assembly quality of metaviromes, compared to short reads alone.
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