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First Report of Prunus virus F Infecting Sweet Cherry ( Prunus avium cv. Staccato) in Canada

2018; American Phytopathological Society; Volume: 102; Issue: 7 Linguagem: Inglês

10.1094/pdis-12-17-1883-pdn

1943-7692

D. James, James Phelan, G.D. Jesperson,

Plant Pathogens and Fungal Diseases

HomePlant DiseaseVol. 102, No. 7First Report of Prunus virus F Infecting Sweet Cherry (Prunus avium cv. Staccato) in Canada PreviousNext DISEASE NOTES OPENOpen Access licenseFirst Report of Prunus virus F Infecting Sweet Cherry (Prunus avium cv. Staccato) in CanadaD. James, J. Phelan, and G. JespersonD. James†Corresponding author: D. James; E-mail: E-mail Address: Delano.James@inspection.gc.ca, J. Phelan, and G. JespersonAffiliationsAuthors and Affiliations D. James † J. Phelan , Canadian Food Inspection Agency, Sidney Laboratory, North Saanich, BC, Canada G. Jesperson , British Columbia Ministry of Agriculture, Kelowna, BC, Canada. Published Online:7 May 2018https://doi.org/10.1094/PDIS-12-17-1883-PDNAboutSections ToolsAdd to favoritesDownload CitationsTrack Citations ShareShare onFacebookTwitterLinked InRedditEmailWechat Villamor et al. (2017) described the first detection in sweet cherry (Prunus avium) of a virus, with a genome organization similar to species in the genus Fabavirus, tentatively named Prunus virus F (PrVF). PrVF was detected in both symptomatic and asymptomatic trees. Sweet cherry cv. Staccato growing in the Okanagan Valley, BC, Canada, and showing a range of symptoms (leaf deformity and some dieback), and asymptomatic trees as controls, were sampled in June 2016. They were screened initially by next-generation sequencing (NGS) followed by RT-PCR testing for confirmation. For NGS analysis, total RNA was extracted initially from two symptomatic plants and one asymptomatic plant using QIAGEN's RNeasy Plant Mini Kit (Ontario, Canada) as described by Kalinowska et al. (2012). For NGS analysis, 500 ng of total RNA was used as the input material and poly(A) RNA was enriched with oligo-dT beads. Illumina TruSeq Stranded mRNA Library Preparation protocol was used to generate the complementary (c) DNA libraries. The RT-PCR tests were: (a) as described by Villamor et al. (2017) targeting RNA-2; and (b) targeting RNA-1 using primers PVF1-CAF (5′-GARAGTTTCCTGARTGGATGCG-3′) and PVF1-CAR (5′-ACACTTGGGCAACATAACTGC-3′) designed in this study amplifying a 457-bp product. RT-PCR conditions were: reverse transcription with denaturation at 94°C for 5 mins followed by 42°C for 50 mins; the conditions for PCR were an initial denaturation step at 94°C for 2 mins followed by 35 cycles of 94°C for 30 s, 55°C for 30 s, and 72°C for 1 min, with a final extension of 72°C for 5 mins. Two 'Staccato' sweet cherry trees were positive by NGS: OKStac-3B/CA1 (symptomatic) and OKStac-4B/CA2 (asymptomatic). NGS analysis generated an average of 90 million reads, with approximately 93,000 and 239,000 mapping to the RNA-1 and RNA-2, respectively, of the CA1 isolate; and approximately 687,000 and 885,000 reads mapping to the RNA-1 and RNA-2, respectively, of the CA2 isolate. Fragments 2,097 bp in size were assembled for RNA-1 (C-terminus of the polymerase coding region). The RNA-2 associated fragments were 1,695 bp in size (CP-L and CP-S coding regions). The sequences are deposited in GenBank: MG459013 and MG459014 for RNA-1 and RNA-2 fragments of the CA1 isolate, respectively; and MG490861 and MG490862 for RNA-1 and RNA-2 fragments of the CA2 isolate, respectively. Compared with PrVF-8816-v1 (KX269865) and PrVF-8816-v2 (KX269866) the CA1 RNA-1 fragment (nt) is 98 and 85% identical, respectively, with aa identities of 100 and 97%, respectively. CA2 RNA-1 identities are 82% (nt) and 94% (aa) identical in both cases. Compared with PrVF-8816-s1 (KX269871) and PrVF-8816-s2 (KX269872), the CA1 RNA-2 fragment is 83% nt (94% aa) and 95% nt (98% aa) identical to the corresponding sequences, respectively. The CA2 RNA-2 fragment is 83% nt (93% aa) and 81% nt (93% aa) identical, respectively. PrVF isolates CA1 and CA2 were detected by RT-PCR (a) amplifying the expected 739-bp fragment. Amplicons were most similar to PrVF-8816-s2 (KX269872) with approximately 97% nt (100% aa) and 82% nt (97% aa) identity. Both isolates were detected by RT-PCR (b), with the expected 457-bp product amplified. PrVF may be widely distributed in cherry as the recent report by Šafářová et al. (2017) described the detection of the virus in sour cherry. More comprehensive information about the distribution of PrVF will help determine biological significance. This is the first report describing the detection of PrVF in cherry in Canada.References:Kalinowska, E., et al. 2012. Acta Biochim. Pol. 59:391. Crossref, ISI, Google ScholarŠafářová, D., et al. 2017. Plant Dis. 101:845. https://doi.org/10.1094/PDIS-09-16-1289-PDN Link, ISI, Google ScholarVillamor, D. E. V., et al. 2017. Arch. Virol. 162:811. https://doi.org/10.1007/s00705-016-3141-z Crossref, ISI, Google ScholarDetailsFiguresLiterature CitedRelated Vol. 102, No. 7 July 2018SubscribeISSN:0191-2917e-ISSN:1943-7692 Metrics Article History Issue Date: 27 Jun 2018Published: 7 May 2018First Look: 18 Jan 2018Accepted: 16 Jan 2018 Page: 1468 InformationThis article is in the public domain and not copyrightable. It may be freely reprinted with customary crediting of the source. The American Phytopathological Society, 2018.Cited byPrunus virus FCABI Compendium, Vol. CABI CompendiumReliable Methodologies and Impactful Tools to Control Fruit Tree Viruses19 June 2021 | Crops, Vol. 1, No. 1Is There a "Biological Desert" With the Discovery of New Plant Viruses? 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