Propriedades biológicas e análise parcial do RNA1 codificador da replicase de isolados do Bean rugose mosaic virus

Detalhes bibliográficos
Autor(a) principal: Barboza, Antonio Augusto Lazarini
Data de Publicação: 2010
Tipo de documento: Tese
Idioma: por
Título da fonte: Repositório Institucional da Universidade Estadual de Maringá (RI-UEM)
Texto Completo: http://repositorio.uem.br:8080/jspui/handle/1/1160
Resumo: Bean rugose mosaic (BRMV) is a member of the Comovirus genus. The virus genome consists of two molecules of single strand RNA, positive sense, called RNA1 and RNA2. BRMV has a restricted host range, causing systemic symptoms such as blistering and leaf deformation on beans. Soybean and bean plants showing characteristic symptoms of BRMV infection were collected in Paraná state. The isolates were maintained in 'Rio Tibagi' bean plants in greenhouse conditions. The virus purified preparation gave three peaks of absorbance readings that could be correlated to the comoviruses particle components. Indirect ELISA tests pointed to replication differential rates for each virus isolate that remained constant even after virus mixed infections. The RNA of both isolates was extracted and a protocol for semi-nested RT-PCR was used to amplify a RNA1 fragment of 700bp, that was cloned and sequenced. Comparisons of the nucleotide sequence with others from the Genbank showed 79% identity with RNA1 of Bean pod mottle virus. The identification of the RNA polymerase coded by BRMV RNA1 was confirmed through the GDD amino acid triad that is conserved in the RNA replicase of all positive sense viruses. The phylogenetic analysis allowed the classification of BRMV as a member of the Secoviridae family. BRMV could be situated next to BPMV, among other comoviruses infecting leguminosae plants. The sequence of the 3' RNA1 region of BRMV is deposited in the GenBank under the accession number GU205154 or gi284807373.
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spelling Propriedades biológicas e análise parcial do RNA1 codificador da replicase de isolados do Bean rugose mosaic virusFeijoeiroSojaDoenças e pragasFeijão - CultivoCiências AgráriasAgronomiaBean rugose mosaic (BRMV) is a member of the Comovirus genus. The virus genome consists of two molecules of single strand RNA, positive sense, called RNA1 and RNA2. BRMV has a restricted host range, causing systemic symptoms such as blistering and leaf deformation on beans. Soybean and bean plants showing characteristic symptoms of BRMV infection were collected in Paraná state. The isolates were maintained in 'Rio Tibagi' bean plants in greenhouse conditions. The virus purified preparation gave three peaks of absorbance readings that could be correlated to the comoviruses particle components. Indirect ELISA tests pointed to replication differential rates for each virus isolate that remained constant even after virus mixed infections. The RNA of both isolates was extracted and a protocol for semi-nested RT-PCR was used to amplify a RNA1 fragment of 700bp, that was cloned and sequenced. Comparisons of the nucleotide sequence with others from the Genbank showed 79% identity with RNA1 of Bean pod mottle virus. The identification of the RNA polymerase coded by BRMV RNA1 was confirmed through the GDD amino acid triad that is conserved in the RNA replicase of all positive sense viruses. The phylogenetic analysis allowed the classification of BRMV as a member of the Secoviridae family. BRMV could be situated next to BPMV, among other comoviruses infecting leguminosae plants. The sequence of the 3' RNA1 region of BRMV is deposited in the GenBank under the accession number GU205154 or gi284807373.O Bean rugose mosaic virus (BRMV) pertence ao gênero Comovirus, possui genoma constituído de duas moléculas de RNA fitas simples, senso positivo, denominadas de RNA1 e RNA2 e apresenta gama de hospedeiras restrita, nas quais induz sintomas sistêmicos de mosaico, bolhosidade e deformação foliar no feijoeiro. Plantas de soja e de feijão apresentando sintomas característicos de infecção por BRMV foram coletadas no Paraná. Os isolados foram mantidos em casa de vegetação em plantas de feijoeiro cv. Rio Tibagi. Os isolados foram purificados, permitindo a constatação da presença de três picos de leitura de absorbância que puderam ser correlacionados aos componentes virais de Comovirus. Ensaios de Elisa indireto apontaram para diferenças na concentração relativa individual de cada isolado, a qual se manteve inalterada mesmo nas infecções mistas. O RNA dos isolados foi extraído e um protocolo de semi-nested RT-PCR foi utilizado na amplificação de um fragmento de 700pb do RNA1, o qual foi clonado e sequenciado. Comparações com outras sequências do Genbank indicaram uma identidade de 79% dessa seqüência com o RNA1 do Bean pod mottle virus. Através da identificação da tríade de aminoácidos GDD, que é conservada nas replicases dos vírus de RNA senso positivo, foi identificada a região conservada da RNA polimerase, codificada pelo RNA1 do BRMV. A análise filogenética permitiu a classificação do BRMV como membro da família Secoviridae, agrupando-o próximo ao BPMV, bem como de outros comovírus de leguminosas. A sequência da região 3' do RNA1 do BRMV encontra-se depositada no GenBank sob o código de acesso GU205154 ou gi284807373.66 fUniversidade Estadual de MaringáBrasilPrograma de Pós-Graduação em AgronomiaUEMMaringá, PREliezer Rodrigues de SoutoCarlos Alberto de Bastos Andrade - UEMClaudete Aparecida Mangolin - UEMÁlvaro Manuel Rodrigues Almeida - EMBRAPAAnésio Bianchini - IAPARBarboza, Antonio Augusto Lazarini2018-04-04T17:26:23Z2018-04-04T17:26:23Z2010info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesishttp://repositorio.uem.br:8080/jspui/handle/1/1160porinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da Universidade Estadual de Maringá (RI-UEM)instname:Universidade Estadual de Maringá (UEM)instacron:UEM2018-10-22T18:07:36Zoai:localhost:1/1160Repositório InstitucionalPUBhttp://repositorio.uem.br:8080/oai/requestopendoar:2024-04-23T14:54:03.895019Repositório Institucional da Universidade Estadual de Maringá (RI-UEM) - Universidade Estadual de Maringá (UEM)false
dc.title.none.fl_str_mv Propriedades biológicas e análise parcial do RNA1 codificador da replicase de isolados do Bean rugose mosaic virus
title Propriedades biológicas e análise parcial do RNA1 codificador da replicase de isolados do Bean rugose mosaic virus
spellingShingle Propriedades biológicas e análise parcial do RNA1 codificador da replicase de isolados do Bean rugose mosaic virus
Barboza, Antonio Augusto Lazarini
Feijoeiro
Soja
Doenças e pragas
Feijão - Cultivo
Ciências Agrárias
Agronomia
title_short Propriedades biológicas e análise parcial do RNA1 codificador da replicase de isolados do Bean rugose mosaic virus
title_full Propriedades biológicas e análise parcial do RNA1 codificador da replicase de isolados do Bean rugose mosaic virus
title_fullStr Propriedades biológicas e análise parcial do RNA1 codificador da replicase de isolados do Bean rugose mosaic virus
title_full_unstemmed Propriedades biológicas e análise parcial do RNA1 codificador da replicase de isolados do Bean rugose mosaic virus
title_sort Propriedades biológicas e análise parcial do RNA1 codificador da replicase de isolados do Bean rugose mosaic virus
author Barboza, Antonio Augusto Lazarini
author_facet Barboza, Antonio Augusto Lazarini
author_role author
dc.contributor.none.fl_str_mv Eliezer Rodrigues de Souto
Carlos Alberto de Bastos Andrade - UEM
Claudete Aparecida Mangolin - UEM
Álvaro Manuel Rodrigues Almeida - EMBRAPA
Anésio Bianchini - IAPAR
dc.contributor.author.fl_str_mv Barboza, Antonio Augusto Lazarini
dc.subject.por.fl_str_mv Feijoeiro
Soja
Doenças e pragas
Feijão - Cultivo
Ciências Agrárias
Agronomia
topic Feijoeiro
Soja
Doenças e pragas
Feijão - Cultivo
Ciências Agrárias
Agronomia
description Bean rugose mosaic (BRMV) is a member of the Comovirus genus. The virus genome consists of two molecules of single strand RNA, positive sense, called RNA1 and RNA2. BRMV has a restricted host range, causing systemic symptoms such as blistering and leaf deformation on beans. Soybean and bean plants showing characteristic symptoms of BRMV infection were collected in Paraná state. The isolates were maintained in 'Rio Tibagi' bean plants in greenhouse conditions. The virus purified preparation gave three peaks of absorbance readings that could be correlated to the comoviruses particle components. Indirect ELISA tests pointed to replication differential rates for each virus isolate that remained constant even after virus mixed infections. The RNA of both isolates was extracted and a protocol for semi-nested RT-PCR was used to amplify a RNA1 fragment of 700bp, that was cloned and sequenced. Comparisons of the nucleotide sequence with others from the Genbank showed 79% identity with RNA1 of Bean pod mottle virus. The identification of the RNA polymerase coded by BRMV RNA1 was confirmed through the GDD amino acid triad that is conserved in the RNA replicase of all positive sense viruses. The phylogenetic analysis allowed the classification of BRMV as a member of the Secoviridae family. BRMV could be situated next to BPMV, among other comoviruses infecting leguminosae plants. The sequence of the 3' RNA1 region of BRMV is deposited in the GenBank under the accession number GU205154 or gi284807373.
publishDate 2010
dc.date.none.fl_str_mv 2010
2018-04-04T17:26:23Z
2018-04-04T17:26:23Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/doctoralThesis
format doctoralThesis
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://repositorio.uem.br:8080/jspui/handle/1/1160
url http://repositorio.uem.br:8080/jspui/handle/1/1160
dc.language.iso.fl_str_mv por
language por
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Universidade Estadual de Maringá
Brasil
Programa de Pós-Graduação em Agronomia
UEM
Maringá, PR
publisher.none.fl_str_mv Universidade Estadual de Maringá
Brasil
Programa de Pós-Graduação em Agronomia
UEM
Maringá, PR
dc.source.none.fl_str_mv reponame:Repositório Institucional da Universidade Estadual de Maringá (RI-UEM)
instname:Universidade Estadual de Maringá (UEM)
instacron:UEM
instname_str Universidade Estadual de Maringá (UEM)
instacron_str UEM
institution UEM
reponame_str Repositório Institucional da Universidade Estadual de Maringá (RI-UEM)
collection Repositório Institucional da Universidade Estadual de Maringá (RI-UEM)
repository.name.fl_str_mv Repositório Institucional da Universidade Estadual de Maringá (RI-UEM) - Universidade Estadual de Maringá (UEM)
repository.mail.fl_str_mv
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