First Report of Pepper Cryptic Virus 2 Infecting Pepper ( Capsicum annum ) in Slovakia
2020; American Phytopathological Society; Volume: 104; Issue: 5 Linguagem: Inglês
10.1094/pdis-12-19-2577-pdn
ISSN1943-7692
AutoresJana Tomašechová, Lukáš Predajňa, Nina Sihelská, Ján Kraic, Daniel Mihálik, Katarína Šoltýs, Miroslav Glasa,
Tópico(s)Plant-Microbe Interactions and Immunity
ResumoHomePlant DiseaseVol. 104, No. 5First Report of Pepper Cryptic Virus 2 Infecting Pepper (Capsicum annum) in Slovakia PreviousNext DISEASE NOTES OPENOpen Access licenseFirst Report of Pepper Cryptic Virus 2 Infecting Pepper (Capsicum annum) in SlovakiaJ. Tomašechová, L. Predajňa, N. Sihelská, J. Kraic, D. Mihálik, K. Šoltys, and M. GlasaJ. TomašechováBiomedical Research Center of the Slovak Academy of Sciences, Institute of Virology, Bratislava, Slovak RepublicUniversity of Ss. Cyril and Methodius, Faculty of Natural Sciences, Trnava, Slovak Republic, L. PredajňaBiomedical Research Center of the Slovak Academy of Sciences, Institute of Virology, Bratislava, Slovak Republic, N. SihelskáBiomedical Research Center of the Slovak Academy of Sciences, Institute of Virology, Bratislava, Slovak Republic, J. KraicUniversity of Ss. Cyril and Methodius, Faculty of Natural Sciences, Trnava, Slovak Republic, D. MihálikUniversity of Ss. Cyril and Methodius, Faculty of Natural Sciences, Trnava, Slovak Republic, K. ŠoltysComenius University Science Park, Comenius University in Bratislava, Bratislava, Slovak RepublicFaculty of Natural Sciences, Comenius University in Bratislava, Bratislava, Slovak Republic, and M. Glasa†Corresponding author: M. Glasa; E-mail Address: [email protected]http://orcid.org/0000-0002-8495-7971Biomedical Research Center of the Slovak Academy of Sciences, Institute of Virology, Bratislava, Slovak RepublicUniversity of Ss. Cyril and Methodius, Faculty of Natural Sciences, Trnava, Slovak RepublicAffiliationsAuthors and Affiliations J. Tomašechová1 2 L. Predajňa1 N. Sihelská1 J. Kraic2 D. Mihálik2 K. Šoltys3 4 M. Glasa1 2 † 1Biomedical Research Center of the Slovak Academy of Sciences, Institute of Virology, Bratislava, Slovak Republic 2University of Ss. Cyril and Methodius, Faculty of Natural Sciences, Trnava, Slovak Republic 3Comenius University Science Park, Comenius University in Bratislava, Bratislava, Slovak Republic 4Faculty of Natural Sciences, Comenius University in Bratislava, Bratislava, Slovak Republic Published Online:8 Mar 2020https://doi.org/10.1094/PDIS-12-19-2577-PDNAboutSectionsSupplemental ToolsAdd to favoritesDownload CitationsTrack Citations ShareShare onFacebookTwitterLinked InRedditEmailWechat In an effort to unravel the virome present in a symptomatic sweet pepper (Capsicum annum) from western Slovakia (locality Čachtice), a plant of cultivar Promotor (coded N65) showing leaf chlorosis, mottling, and deformation was sampled in August 2017 and subjected to high-throughput sequencing (HTS). Total RNAs were isolated from fully developed leaves of the plant using the Spectrum Plant Total RNA Kit (Sigma Aldrich, St. Louis, MO). Ribosomal RNA-depleted RNA preparations were used for cDNA library synthesis (Nextera XT, Illumina, San Diego, CA). HTS was carried out on an Illumina MiSeq platform (300-bp paired-end sequencing), and the obtained high-quality reads (∼5.5 million) were analyzed using Geneious software (version 8). BLASTn analysis of the generated contigs (∼16,000 contigs >500 bp) revealed a complex viral infection involving cucumber mosaic virus (CMV), watermelon mosaic virus, bell pepper endornavirus, and pepper cryptic virus 2 (PCV-2) (Tomašechová et al. 2020). Two de novo assembled contigs (1,574 and 1,493 bp in length) corresponded to nearly complete PCV-2 RNA1 and RNA2 (genome coverage 98.3 and 99.7%), with respectively 98.9 and 99.5% nucleotide identity to the references NC_034159 and NC_034167 of PCV-2. The sequences, which have been deposited in GenBank under the accession numbers MN723540 and MN723541, showed close phylogenetic relationship with the Turkish Tur isolate, having mutual 99.4 and 99.8% identity, respectively. Because PCV-2 was previously unreported from the central European region, to confirm its presence in the N65 plant and to assess further the virus presence in Slovakia, 35 leaf samples obtained from various symptomatic sweet pepper and chili plants were tested in 2018 and 2019 by reverse transcription PCR (RT-PCR) using two newly designed primer pairs: 2PCV1_639F (5′-ACACCCGCACACAATTAACG-3′)/2PCV1_1176R (5′-CCGATTAGTGAATCATCTCC-3′) for RNA1 and 2PCV2_858F (5′-AGGCAACTACTCTATGGAGG-3′)/2PCV2_1272R (5′-CCTATCTGCTTCTCCTGTAG-3′) for RNA2. The expected specific 537- and 414-bp products were amplified from 15 of the 35 samples tested and also from the original N65 pepper plant. The specificity of RT-PCR products was ascertained by Sanger sequencing (accession nos. MN792889 to MN792916). BLASTn analysis indicated that all the Slovak PCV-2 isolates shared high nucleotide identity (97 to 100%) with the corresponding portions of N65 isolate, resulting in low average nucleotide divergence (0.1% for RNA1 and 0.5% for RNA2). Fifteen PCV-2-infected pepper and chili samples were further tested for the presence of two main common pepper viruses in Slovakia: CMV and potato virus Y (PVY) (Glasa, unpublished results). Both RT-PCR (described in Glasa et al. 2019) and double antibody sandwich ELISA tests using commercial antibodies (Bioreba AG, Reinach, Switzerland) revealed mixed PCV-2/PVY (n = 9), PCV-2/CMV (n = 2), or PCV-2/PVY/CMV (n = 4) infections. As a consequence, the symptoms observed (mosaics, leaf mottling, and deformations) could not be attributed to a particular virus or to synergism between these viruses. PCV-2 is a bipartite double-stranded RNA virus belonging to the genus Deltapartitivirus (family Partitiviridae) (Sabanadzovic and Valverde 2011). The virus was previously reported from the United States, India, China, and Turkey (Sabanadzovic and Valverde 2011; Safari and Roossinck 2018; Saritha et al. 2016; Sun et al. 2017). To our knowledge, this is the first report of PCV-2 infection of pepper in Slovakia, further emphasizing a frequent occurrence of persistent viruses in agricultural crops (Roossinck 2015; Safari and Roossinck 2018). Although commonly accepted as causing no symptoms, the contribution of such viruses to the biology of their hosts, often infected by a complex of viruses, remains unclear.The author(s) declare no conflict of interest.References:Glasa, M., et al. 2019. Plant Prot. Sci. 55:159. https://doi.org/10.17221/144/2018-PPS Crossref, ISI, Google ScholarRoossinck, M. J. 2015. Front. Microbiol. 5:767. https://doi.org/10.3389/fmicb.2014.00767 Crossref, ISI, Google ScholarSabanadzovic, S., and Valverde, R. A. 2011. Virus Genes 43:307. https://doi.org/10.1007/s11262-011-0634-4 Crossref, ISI, Google ScholarSafari, M., and Roossinck, M. J. 2018. Mol. Plant-Microbe Interact. 31:766. https://doi.org/10.1094/MPMI-12-17-0312-R Link, ISI, Google ScholarSaritha, R. K., et al. 2016. VirusDis. 27:327. https://doi.org/10.1007/s13337-016-0327-7 Crossref, Google ScholarSun, M., et al. 2017. J. Plant Pathol. 99:298. https://doi.org/10.4454/jpp.v99i1.3808 Google ScholarTomašechová, J., et al. 2020. Plants 9:41. https://doi.org/10.3390/plants9010041 Crossref, Google ScholarThe author(s) declare no conflict of interest.Funding: Funding was provided by European Regional Development Fund (REVOGENE [ITMS: 26240220067]) and Agentúra na Podporu Výskumu a Vývoja (APVV-18-0005).DetailsFiguresLiterature CitedRelated Vol. 104, No. 5 May 2020SubscribeISSN:0191-2917e-ISSN:1943-7692 DownloadCaptionSymptoms observed in the field on zucchini plants caused by Fusarium solani f. sp. cucurbitae (A. Pérez-Hernández et al.). Photo credit: J. M. Gómez-Vázquez. Peach tree with excavated root collar (S. B. Miller et al.). Photo credit: G. Schnabel. Metrics Article History Issue Date: 3 May 2020Published: 8 Mar 2020First Look: 15 Jan 2020Accepted: 12 Jan 2020 Page: 1565 Information© 2020 The American Phytopathological SocietyFundingEuropean Regional Development FundGrant/Award Number: REVOGENE [ITMS: 26240220067]Agentúra na Podporu Výskumu a VývojaGrant/Award Number: APVV-18-0005Keywordsviruses and viroidspathogen diversityfield cropsoilseeds and legumespathogen detectionThe author(s) declare no conflict of interest.Cited byCucumber mosaic virus (cucumber mosaic)CABI Compendium, Vol. CABI CompendiumPepper cryptic virus 2CABI Compendium, Vol. CABI CompendiumWatermelon mosaic virus (watermelon mosaic)CABI Compendium, Vol. CABI CompendiumPotato virus Y (potato mottle)CABI Compendium, Vol. CABI Compendium
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