High-throughput sequencing applied for the identification of viruses infecting grapevines in Brazil and genetic variability analysis.

The application of high-throughput sequencing technologies (HTS) enables the recovery of many nucleotide sequence fragments from diseased plants and may help in pathogen identification. This study was designed to identify viruses infecting 15 grapevine (Vitis spp.) samples collected from experimental fields and vine collections and assess the genetic variability of the identified viruses. The virus-enriched dsRNAs were extracted from bark scrapings and sequenced using an Illumina platform. The paired-end reads were analyzed, assembled contigs were generated and identified as related to viruses. Contigs of 14 viruses have been identified, some of them covering large extensions of viral genomes or resulting in assembly of near-complete or complete genomes. Grapevine virus infections are usually mixed and the HTS assays were suitable to identify ten viruses already reported that traditionally infect grapevines in Brazil, one that has been recently identified (Grapevine Syrah virus 1) and others (Grapevine Cabernet Sauvignon reovirus, Grapevine Red Globe virus and Grapevine vein clearing virus) not previously reported in this country. Nucleotide identities among Brazilian isolates identified by HTS and homologous grapevine virus sequences in GenBank were high, ranging from 77% to 99%. Genetic variability analysis of viral sequences obtained by HTS and sequences available in GenBank indicated that the coding regions in the different viral species are under purifying selection, and that recombination events occurred in the majority of the viral species analyzed. The coat protein genes, generally, had lower genetic variability than the replicase and movement protein genes. Keywords Vitis . Diagnosis . HTS . Next-generation sequencing . NGS . Variability

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Bibliographic Details
Main Authors: FAJARDO, T. V. M., SILVA, F. N., EIRAS, M., NICKEL, O.
Other Authors: THOR VINICIUS MARTINS FAJARDO, CNPUV; Fábio N. Silva, Universidade do Estado de Santa Catarina, Lages, SC 88520-000, Brazil; Marcelo Eiras, Instituto Biológico de São Paulo, São Paulo, SP 04014-002, Brazil; OSMAR NICKEL, CNPUV.
Format: Artigo de periódico biblioteca
Language:pt_BR
por
Published: 2017-10-23
Subjects:Diagnosis, HTS, Next-generation sequencing, NGS, Vitis, variability,
Online Access:http://www.alice.cnptia.embrapa.br/alice/handle/doc/1077833
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spelling dig-alice-doc-10778332019-05-07T00:44:17Z High-throughput sequencing applied for the identification of viruses infecting grapevines in Brazil and genetic variability analysis. FAJARDO, T. V. M. SILVA, F. N. EIRAS, M. NICKEL, O. THOR VINICIUS MARTINS FAJARDO, CNPUV; Fábio N. Silva, Universidade do Estado de Santa Catarina, Lages, SC 88520-000, Brazil; Marcelo Eiras, Instituto Biológico de São Paulo, São Paulo, SP 04014-002, Brazil; OSMAR NICKEL, CNPUV. Diagnosis HTS Next-generation sequencing NGS Vitis variability The application of high-throughput sequencing technologies (HTS) enables the recovery of many nucleotide sequence fragments from diseased plants and may help in pathogen identification. This study was designed to identify viruses infecting 15 grapevine (Vitis spp.) samples collected from experimental fields and vine collections and assess the genetic variability of the identified viruses. The virus-enriched dsRNAs were extracted from bark scrapings and sequenced using an Illumina platform. The paired-end reads were analyzed, assembled contigs were generated and identified as related to viruses. Contigs of 14 viruses have been identified, some of them covering large extensions of viral genomes or resulting in assembly of near-complete or complete genomes. Grapevine virus infections are usually mixed and the HTS assays were suitable to identify ten viruses already reported that traditionally infect grapevines in Brazil, one that has been recently identified (Grapevine Syrah virus 1) and others (Grapevine Cabernet Sauvignon reovirus, Grapevine Red Globe virus and Grapevine vein clearing virus) not previously reported in this country. Nucleotide identities among Brazilian isolates identified by HTS and homologous grapevine virus sequences in GenBank were high, ranging from 77% to 99%. Genetic variability analysis of viral sequences obtained by HTS and sequences available in GenBank indicated that the coding regions in the different viral species are under purifying selection, and that recombination events occurred in the majority of the viral species analyzed. The coat protein genes, generally, had lower genetic variability than the replicase and movement protein genes. Keywords Vitis . Diagnosis . HTS . Next-generation sequencing . NGS . Variability 2019-05-07T00:44:09Z 2019-05-07T00:44:09Z 2017-10-23 2017 2019-05-07T00:44:09Z Artigo de periódico Tropical Plant Pathology, v. 52, p. 250-260, 2017. http://www.alice.cnptia.embrapa.br/alice/handle/doc/1077833 10.1007/s40858-017-0142-8 pt_BR por openAccess
institution EMBRAPA
collection DSpace
country Brasil
countrycode BR
component Bibliográfico
access En linea
databasecode dig-alice
tag biblioteca
region America del Sur
libraryname Sistema de bibliotecas de EMBRAPA
language pt_BR
por
topic Diagnosis
HTS
Next-generation sequencing
NGS
Vitis
variability
Diagnosis
HTS
Next-generation sequencing
NGS
Vitis
variability
spellingShingle Diagnosis
HTS
Next-generation sequencing
NGS
Vitis
variability
Diagnosis
HTS
Next-generation sequencing
NGS
Vitis
variability
FAJARDO, T. V. M.
SILVA, F. N.
EIRAS, M.
NICKEL, O.
High-throughput sequencing applied for the identification of viruses infecting grapevines in Brazil and genetic variability analysis.
description The application of high-throughput sequencing technologies (HTS) enables the recovery of many nucleotide sequence fragments from diseased plants and may help in pathogen identification. This study was designed to identify viruses infecting 15 grapevine (Vitis spp.) samples collected from experimental fields and vine collections and assess the genetic variability of the identified viruses. The virus-enriched dsRNAs were extracted from bark scrapings and sequenced using an Illumina platform. The paired-end reads were analyzed, assembled contigs were generated and identified as related to viruses. Contigs of 14 viruses have been identified, some of them covering large extensions of viral genomes or resulting in assembly of near-complete or complete genomes. Grapevine virus infections are usually mixed and the HTS assays were suitable to identify ten viruses already reported that traditionally infect grapevines in Brazil, one that has been recently identified (Grapevine Syrah virus 1) and others (Grapevine Cabernet Sauvignon reovirus, Grapevine Red Globe virus and Grapevine vein clearing virus) not previously reported in this country. Nucleotide identities among Brazilian isolates identified by HTS and homologous grapevine virus sequences in GenBank were high, ranging from 77% to 99%. Genetic variability analysis of viral sequences obtained by HTS and sequences available in GenBank indicated that the coding regions in the different viral species are under purifying selection, and that recombination events occurred in the majority of the viral species analyzed. The coat protein genes, generally, had lower genetic variability than the replicase and movement protein genes. Keywords Vitis . Diagnosis . HTS . Next-generation sequencing . NGS . Variability
author2 THOR VINICIUS MARTINS FAJARDO, CNPUV; Fábio N. Silva, Universidade do Estado de Santa Catarina, Lages, SC 88520-000, Brazil; Marcelo Eiras, Instituto Biológico de São Paulo, São Paulo, SP 04014-002, Brazil; OSMAR NICKEL, CNPUV.
author_facet THOR VINICIUS MARTINS FAJARDO, CNPUV; Fábio N. Silva, Universidade do Estado de Santa Catarina, Lages, SC 88520-000, Brazil; Marcelo Eiras, Instituto Biológico de São Paulo, São Paulo, SP 04014-002, Brazil; OSMAR NICKEL, CNPUV.
FAJARDO, T. V. M.
SILVA, F. N.
EIRAS, M.
NICKEL, O.
format Artigo de periódico
topic_facet Diagnosis
HTS
Next-generation sequencing
NGS
Vitis
variability
author FAJARDO, T. V. M.
SILVA, F. N.
EIRAS, M.
NICKEL, O.
author_sort FAJARDO, T. V. M.
title High-throughput sequencing applied for the identification of viruses infecting grapevines in Brazil and genetic variability analysis.
title_short High-throughput sequencing applied for the identification of viruses infecting grapevines in Brazil and genetic variability analysis.
title_full High-throughput sequencing applied for the identification of viruses infecting grapevines in Brazil and genetic variability analysis.
title_fullStr High-throughput sequencing applied for the identification of viruses infecting grapevines in Brazil and genetic variability analysis.
title_full_unstemmed High-throughput sequencing applied for the identification of viruses infecting grapevines in Brazil and genetic variability analysis.
title_sort high-throughput sequencing applied for the identification of viruses infecting grapevines in brazil and genetic variability analysis.
publishDate 2017-10-23
url http://www.alice.cnptia.embrapa.br/alice/handle/doc/1077833
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