Grapevines (Vitis sp.) can host up to 86 virus species, some of which affect plant vigor, production and fruit quality. In 2019, cv. BRS Bibiana, a hybrid vine showing mild leafroll symptoms, collected in an experimental field in Canoinhas-SC, Brazil, was investigated for viruses.Evento online

FAJARDO, T. V. M.

NHANI JUNIOR, A.

NICKEL, O.

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194  ANÁLISE DO VIROMA POR HTS REVELA INFECÇÃO MÚLTIPLA EM PLANTA DE VIDEIRA ANALYSIS OF VIROME BY HIGH-THROUGHPUT SEQUENCING REVEALED MULTIPLE INFECTION IN A SINGLE GRAPEVINE PLANT  THOR VINÍCIUS MARTINS FAJARDO 1; ANTONIO NHANI JÚNIOR 2; OSMAR NICKEL 3 1Pesquisador. Bento Gonçalves-RS, Brasil. Embrapa Uva e Vinho; 2Pesquisador. Campinas-SP, Brasil. Embrapa Informática Agropecuária; 3Pesquisador. Bento Gonçalves-RS, Brasil. Embrapa Uva e Vinho  Resumo:  Grapevines (Vitis sp.) can host up to 86 virus species, some of which affect plant vigor, production and fruit quality. In 2019, cv. BRS Bibiana, a hybrid vine showing mild leafroll symptoms, collected in an experimental field in Canoinhas-SC, Brazil, was investigated for viruses. Ribosomal-depleted total RNA (TNA) from this sample was used to prepare a sequencing library generated with TruSeq Stranded Total RNA with Ribo-Zero Plant kit (Illumina, USA), then subjected to high-throughput sequencing (HTS) on Illumina HiSeq X Ten platform. The HTS yielded 45,419,722 total reads, which were quality trimmed with FastQ and Trim Galore softwares. De novo assembled contigs built by SPAdes (rnaSPAdes) software generated 256,894 contigs, some of which, after QUAST (MetaQUAST) analysis, corresponded to virus/viroid genomes that were identified by comparison with viral/viroidal reference sequences available in the NCBI database. In total, four RNA viruses/viroids, common grapevine pathogens, were identified in the BlastN/BlastX analyses. The following obtained sequences were deposited in GenBank: grapevine rupestris stem pitting-associated virus (GRSPaV, Foveavirus, Betaflexiviridae), GRSPaV-BIB-BR isolate, 8,679 nucleotides (nt), GenBank accession code MW357717; grapevine virus B (GVB, Vitivirus, Betaflexiviridae), GVB-BIB-BR isolate, 7,599 nt, MW357718; grapevine yellow speckle viroid 1 (GYSVd-1, Apscaviroid, Pospiviroidae), GYSVd1-BIB-BR isolate, 366 nt, MW357719, and hop stunt viroid (HSVd, Hostuviroid, Pospiviroidae), HSVd-BIB-BR isolate, 297 nt, MW357720. The highest pairwise nt sequence identities between these Brazilian and homologous isolates (with query cover 99-100%) showed 99.39% nt identity with Brazilian GRSPaV NUB1-BR isolate (MK804766); 97.81% nt identity with Brazilian GVB ISA-BR isolate (KX790785); 99.73% nt identity with Nigerian GYSVd1 UDSV1-4 isolate (MF576400), and 100% nt identity with Canadian and Chinese HSVd BacSB11_5310, BacMF3_243, BacMF3_783, and chi100Y-02-4-5 isolates (MT769774, MT769768, MT769769, AB219944), indicating high homologies among compared isolates. To confirm the HTS results, fresh TNA extract from original source was screened by conventional or RT-qPCR using specific primers and/or probes. The results demonstrate the complexity of grapevine viral diseases and show that mixed virus/viroid infection of grapevine can be considered the rule, since this condition represents the majority of cases. Palavras-chave: Vine; HTS; NGS; Virus; Viroid Apoio  To Empresa Brasileira de Pesquisa Agropecuária (Embrapa) for the financial support to this research.  

Analysis of virome by high-throughput sequencing revealed multiple infection in a single grapevine plant.

http://www.alice.cnptia.embrapa.br/bitstream/doc/1134786/1/Thor-2021-p194-anais-cbfito.pdf

Analysis of virome by high-throughput sequencing revealed multiple infection in a single grapevine plant.

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