dc.contributor.author |
Blouin, Arnaud |
en |
dc.contributor.author |
Ross, Howard |
en |
dc.contributor.author |
Hobson-Peters, J |
en |
dc.contributor.author |
O'Brien, CA |
en |
dc.contributor.author |
Warren, B |
en |
dc.contributor.author |
MacDiarmid, Robin |
en |
dc.date.accessioned |
2016-11-28T03:04:58Z |
en |
dc.date.issued |
2016-09 |
en |
dc.identifier.citation |
Molecular Ecology Resources 16(5):1255-1263 Sep 2016 |
en |
dc.identifier.issn |
1755-098X |
en |
dc.identifier.uri |
http://hdl.handle.net/2292/31161 |
en |
dc.description.abstract |
Next-generation sequencing technologies enable the rapid identification of viral infection of diseased organisms. However, despite a consistent decrease in sequencing costs, it is difficult to justify their use in large-scale surveys without a virus sequence enrichment technique. As the majority of plant viruses have an RNA genome, a common approach is to extract the double-stranded RNA (dsRNA) replicative form, to enrich the replicating virus genetic material over the host background. The traditional dsRNA extraction is time-consuming and labour-intensive. We present an alternative method to enrich dsRNA from plant extracts using anti-dsRNA monoclonal antibodies in a pull-down assay. The extracted dsRNA can be amplified by reverse transcriptase–polymerase chain reaction and sequenced by next-generation sequencing. In our study, we have selected three distinct plant hosts: Māori potato (Solanum tuberosum), rengarenga (Arthropodium cirratum) and broadleaved dock (Rumex obtusifolius) representing a cultivated crop, a New Zealand-native ornamental plant and a weed, respectively. Of the sequence data obtained, 31–74% of the reads were of viral origin, and we identified five viruses including Potato virus Y and Potato virus S in potato; Turnip mosaic virus in rengarenga (a new host record); and in the dock sample Cherry leaf roll virus and a novel virus belonging to the genus Macluravirus. We believe that this new assay represents a significant opportunity to upscale virus ecology studies from environmental, primary industry and/or medical samples. |
en |
dc.relation.ispartofseries |
Molecular Ecology Resources |
en |
dc.rights |
Items in ResearchSpace are protected by copyright, with all rights reserved, unless otherwise indicated. Previously published items are made available in accordance with the copyright policy of the publisher. |
en |
dc.rights.uri |
https://researchspace.auckland.ac.nz/docs/uoa-docs/rights.htm |
en |
dc.title |
A new virus discovered by immunocapture of double-stranded RNA, a rapid method for virus enrichment in metagenomic studies |
en |
dc.type |
Journal Article |
en |
dc.identifier.doi |
10.1111/1755-0998.12525 |
en |
pubs.issue |
5 |
en |
pubs.begin-page |
1255 |
en |
pubs.volume |
16 |
en |
dc.identifier.pmid |
26990372 |
en |
pubs.end-page |
1263 |
en |
pubs.publication-status |
Accepted |
en |
dc.rights.accessrights |
http://purl.org/eprint/accessRights/RestrictedAccess |
en |
pubs.subtype |
Article |
en |
pubs.elements-id |
520563 |
en |
pubs.org-id |
Science |
en |
pubs.org-id |
Biological Sciences |
en |
dc.identifier.eissn |
1755-0998 |
en |
pubs.record-created-at-source-date |
2016-02-09 |
en |
pubs.dimensions-id |
26990372 |
en |