| Literature DB >> 34064231 |
Katrine Wacenius Skov Alanin1,2, Laura Milena Forero Junco1, Jacob Bruun Jørgensen1, Tue Kjærgaard Nielsen1, Morten Arendt Rasmussen3,4, Witold Kot1, Lars Hestbjerg Hansen1.
Abstract
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 (p = 0.015) of the 47 identified putative Pseudomonas phages with a minimum twofold change above zero in read abundance when adding a Pseudomonas syringae DC3000 host. Surprisingly, it appears that only two out of five plaque-forming phages from the same organic waste sample, targeting the Pseudomonas 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.Entities:
Keywords: Illumina sequencing; Oxford Nanopore Technologies; PEG precipitation; Pseudomonas syringae pv. tomato DC3000; bacteriophages; metaviromes; phage enrichment; plaque-assay; viromes
Year: 2021 PMID: 34064231 DOI: 10.3390/v13060959
Source DB: PubMed Journal: Viruses ISSN: 1999-4915 Impact factor: 5.048