Data from: A new plant virus discovered by immunocapture of double stranded RNA; assessment of a novel approach for viral metagenomics studies
Data files
Mar 10, 2016 version files 13.46 MB
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                CLRV_R1.bam
                20.67 KB
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                CLRV_R2.bam
                9.20 KB
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                PVSA_R1.bam
                1.69 MB
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                PVSA_R2.bam
                732.53 KB
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                PVSO_R1.bam
                2.54 MB
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                PVSO_R2.bam
                1.03 MB
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                PVY_R1.bam
                142.86 KB
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                PVY_R2.bam
                74.88 KB
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                Rumex_R1.bam
                2.83 MB
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                Rumex_R2.bam
                969.37 KB
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                TuMV_R1.bam
                2.08 MB
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                TuMV_R2.bam
                1.33 MB
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.
  
  
  
  