Occurrence of Coleus blumei Viroid 6 in Commercial Coleus blumei in Canada: The First Report Outside of China
2018; American Phytopathological Society; Volume: 103; Issue: 4 Linguagem: Inglês
10.1094/pdis-10-18-1875-pdn
ISSN1943-7692
AutoresR. L. Smith, Janice Lawrence, Malay Shukla, Mathuresh Singh, Xiang Li, Huimin Xu, D. Chen, Kyle M. Gardner, Xianzhou Nie,
Tópico(s)Viral Infectious Diseases and Gene Expression in Insects
ResumoHomePlant DiseaseVol. 103, No. 4Occurrence of Coleus blumei Viroid 6 in Commercial Coleus blumei in Canada: The First Report Outside of China Previous DISEASE NOTES OPENOpen Access licenseOccurrence of Coleus blumei Viroid 6 in Commercial Coleus blumei in Canada: The First Report Outside of ChinaR. L. Smith, J. Lawrence, M. Shukla, M. Singh, X. Li, H. Xu, D. Chen, K. Gardner, and X. NieR. L. SmithFredericton Research and Development Centre, Agriculture and Agri-Food Canada, Fredericton, NB, E3B 4Z7, Canada; Department of Biology, University of New Brunswick, Fredericton, NB, E3B 5A3, Canada; , J. LawrenceDepartment of Biology, University of New Brunswick, Fredericton, NB, E3B 5A3, Canada; , M. ShuklaFredericton Research and Development Centre, Agriculture and Agri-Food Canada, Fredericton, NB, E3B 4Z7, Canada; , M. SinghAgricultural Certification Services, Fredericton, NB, E3B 8B7, Canada; and , X. LiCanadian Food Inspection Agency, Charlottetown Laboratory, Charlottetown, PE, C1A 5T1, Canada, H. XuCanadian Food Inspection Agency, Charlottetown Laboratory, Charlottetown, PE, C1A 5T1, Canada, D. ChenFredericton Research and Development Centre, Agriculture and Agri-Food Canada, Fredericton, NB, E3B 4Z7, Canada; , K. GardnerFredericton Research and Development Centre, Agriculture and Agri-Food Canada, Fredericton, NB, E3B 4Z7, Canada; , and X. Nie†Corresponding author: X. Nie; E-mail: E-mail Address: xianzhou.nie@canada.cahttp://orcid.org/0000-0002-6831-2639Fredericton Research and Development Centre, Agriculture and Agri-Food Canada, Fredericton, NB, E3B 4Z7, Canada; AffiliationsAuthors and Affiliations R. L. Smith1 2 J. Lawrence2 M. Shukla1 M. Singh3 X. Li4 H. Xu4 D. Chen1 K. Gardner1 X. Nie1 † 1Fredericton Research and Development Centre, Agriculture and Agri-Food Canada, Fredericton, NB, E3B 4Z7, Canada; 2Department of Biology, University of New Brunswick, Fredericton, NB, E3B 5A3, Canada; 3Agricultural Certification Services, Fredericton, NB, E3B 8B7, Canada; and 4Canadian Food Inspection Agency, Charlottetown Laboratory, Charlottetown, PE, C1A 5T1, Canada Published Online:14 Feb 2019https://doi.org/10.1094/PDIS-10-18-1875-PDNAboutSectionsSupplemental ToolsAdd to favoritesDownload CitationsTrack Citations ShareShare onFacebookTwitterLinked InRedditEmailWechat The viroid genus Coleviroid (vernacular name: coleviroids), within the family Pospiviroidae, encompasses all viroids discovered to date that infect Plectranthus scutellarioides (synonym: Coleus blumei; vernacular name: coleus), an ornamental plant commonly grown worldwide. This genus includes six viroids: Coleus blumei viroid 1 through 6 (CbVd-1 to 6). Previously, CbVd-1 and CbVd-5 were detected in coleus plants, collected at a local nursery in Fredericton, New Brunswick, Canada (Smith et al. 2018), via Sanger sequencing of cloned amplicons generated from the universal coleviroid primers CbVds-P1 (5′-GCAGCGCTGCAACGGAAT-3′) and CbVds-P2 (5′-GCAGCGCTGCCAGGGAACCCAGGT-3′) (Jiang et al. 2011). However, the low-throughput nature of the amplicon clone-Sanger sequencing approach allowed for the possibility that other coleviroids and sequence variants may have escaped detection. To mitigate this, next generation sequencing (NGS) was performed on amplicons of ∼250 to 350 bp (using the universal coleviroid primer pair) of pooled leaf samples collected at the local nursery in both the summers of 2017 (nine cultivars: Black Dragon, Main Street Michigan Avenue, Pineapple Surprise, Premium Sun Chocolate Covered Cherry, Stained Glassworks Luminesce, Stained Glassworks Raspberry Tart, Stained Glassworks Tilt-a-Whirl, Wizard Coral Sunrise, and Wizard Rose) and 2018 (eight cultivars: Main Street Abbey Road, Main Street Chartres Street, Main Street Fifth Avenue, Main Street Rodeo Drive, Main Street River Walk, Main Street Wall Street, Great Falls Niagara, and Great Falls Yosemite). Sequencing libraries were prepared using the TruSeq Nano DNA LT Library Preparation Kit (Illumina) with a slight modification by employing 100 ng of cDNA as input into step three of the workflow. NGS was performed with a MiSeq system using the MiSeq Reagent Nano Kit version 2 (500 cycles; Illumina). Sequence reads were quality trimmed using Trimmomatic (Bolger et al. 2014) and assembled using Trinity transcriptome assembly pipeline (Grabherr et al. 2011). To determine the presence of specific viroids, unique assembled transcripts were compared with the NCBI nr/nt database using MegaBLAST and also directly aligned to the known genome sequences of CbVd-1 to 6 using Burrows-Wheeler Aligner (Li and Durbin 2009). In addition to CbVd-1 and CbVd-5, NGS detected the presence of CbVd-6 at the nursery in both years of collection. The 342-nt-long CbVd-6 sequence (accession no. MK096271) exhibited 99% identity with the originally reported sequence (Hou et al. 2009), showing a single substitution of G for A at position 331. This sequence was confirmed via Sanger sequencing of plasmid clones inserted with the aforementioned amplicon after gel purification of the region ∼350 bp. To further investigate the presence of CbVd-6, the 17 coleus cultivars from 2017 and 2018 (each a pooling of 10 to 15 leaves from individual plants within the cultivar), and 180 coleus seed plants (attainment and growth described in Smith et al. [2018]), were tested by reverse transcription polymerase chain reaction using, first, the universal coleviroid primer pair, and, if positive for a band ∼350 bp, a newly designed CbVd-6-specific primer pair, CbVd5_F2 (5′-AATAGAGCTTCGCTCGCGAAC-3′) and CbVd6_R (5′-ATGATCCGAGGATCGCTTTCC-3′) (amplicon size 189 bp). In all, CbVd-6 was detected in one cultivar (Stained Glassworks Raspberry Tart) from the year 2017 and two cultivars (Great Falls Niagara and Great Falls Yosemite) from the year 2018. Within the two CbVd-6-positive pooled samples/cultivars analyzed in 2018, all individual plants tested positive for CbVd-6 infection. For the 180 seed plants that had previously been tested for CbVd-1 and CbVd-5 (Smith et al. 2018), none tested positive for CbVd-6. To our knowledge, this is the first detection of CbVd-6 outside of China since its initial description in 2009 (Hou et al. 2009). This finding, together with the previous report of CbVd-5 in Canada, provides evidence that the occurrence of the recently discovered coleviroids (i.e., CbVd-5 and CbVd-6) is not limited to coleus in China only, but instead they could be widespread in coleus plants in other countries. Although coleviroid infections appear to be predominantly asymptomatic in coleus plants (Nie and Singh 2017), much remains unknown about other effects on coleus plants, their host range, and potential risk to other ornamental or horticultural plant species; therefore, precautions may be necessary to minimize the spread of these viroids.References:Bolger, A. M., et al. 2014. 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Plant Dis. 102:1862. https://doi.org/10.1094/PDIS-01-18-0055-PDN Link, ISI, Google ScholarFunding: Funding was provided by Canadian Food Inspection Agency (CHA-P-1601).DetailsFiguresLiterature CitedRelated Vol. 103, No. 4 April 2019SubscribeISSN:0191-2917e-ISSN:1943-7692 DownloadCaptionGray mold on kiwifruit leaves caused by Botrytis cinerea (courtesy Guoshu Gong and Qinjun Tao); sunflower rust on bracts (courtesy Sam Markell); cucumber plant with mosaic symptoms caused by papaya ringspot virus (courtesy Roger Jones). Metrics Article History Issue Date: 10 Apr 2019Published: 14 Feb 2019First Look: 25 Nov 2018Accepted: 24 Nov 2018 Page: 782 Information© 2019 The American Phytopathological SocietyFundingCanadian Food Inspection AgencyGrant/Award Number: CHA-P-1601Cited byColeus blumei viroid 7: a novel viroid resulting from genome recombination between Coleus blumei viroids 1 and 515 August 2021 | Archives of Virology, Vol. 166, No. 11Molecular characterization of Coleus blumei viroids 1 and 3 in Plectranthus scutellarioides in Croatia13 July 2019 | European Journal of Plant Pathology, Vol. 155, No. 3
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