Immunocapture-RT-PCR detection of Cassava common mosaic virus in cassava obtained from meristem-tip culture in Paraná state

Detalhes bibliográficos
Autor(a) principal: Silva,Jaqueline M.
Data de Publicação: 2011
Outros Autores: Carnelossi,Patrícia R., Bijora,Taise, Facco,Cassiele U., Picoli,Marcelo H.S., Souto,Eliezer R., Oliveira,Arildo J.B., Almeida,Álvaro M.R.
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Tropical plant pathology (Online)
Texto Completo: http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1982-56762011000500001
Resumo: A virus survey conducted in the northwest region of Paraná, the main cassava-producing region of that state, showed Cassava common mosaic virus (CsCMV) to be widespread, infecting more than 90% of plants from all cassava cultivars. A CsCMV isolate was purified and used to raise a high-titer (1/1.000) polyclonal antiserum for indexing plants produced from meristem-tip culture, using PTA-ELISA. From an initial production of 110 plants of cultivar Olho Junto, 31 remained infected as indicated by PTA-ELISA. To improve the sensitivity of virus detection, an immunocapture-RT-PCR (IC-RT-PCR) protocol was established. Virus-specific IgG, purified and combined with a primer set for the genus Potexvirus, could readily detect CsCMV in cassava crude leaf extracts. IC-RT-PCR products of 750 bp were amplified from six out of 35 plants previously tested as virus-negative by PTA-ELISA. Sequence analysis of cloned IC-RT-PCR products confirmed they were part of the CsCMV replicase gene, indicating that the virus was still present after thermotherapy and meristem-tip culture. PTA-ELISA enabled initial screening of virus-positive cassava, reducing the number of plants to be further analyzed by IC-RT-PCR. Though CsCMV has been considered of minor importance, its widespread nature, as noticed in Paraná, indicates the need for adoption of effective control measures.
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spelling Immunocapture-RT-PCR detection of Cassava common mosaic virus in cassava obtained from meristem-tip culture in Paraná statePotexvirusEuphorbiaceaeIC-RT-PCRindexingPTA-ELISAA virus survey conducted in the northwest region of Paraná, the main cassava-producing region of that state, showed Cassava common mosaic virus (CsCMV) to be widespread, infecting more than 90% of plants from all cassava cultivars. A CsCMV isolate was purified and used to raise a high-titer (1/1.000) polyclonal antiserum for indexing plants produced from meristem-tip culture, using PTA-ELISA. From an initial production of 110 plants of cultivar Olho Junto, 31 remained infected as indicated by PTA-ELISA. To improve the sensitivity of virus detection, an immunocapture-RT-PCR (IC-RT-PCR) protocol was established. Virus-specific IgG, purified and combined with a primer set for the genus Potexvirus, could readily detect CsCMV in cassava crude leaf extracts. IC-RT-PCR products of 750 bp were amplified from six out of 35 plants previously tested as virus-negative by PTA-ELISA. Sequence analysis of cloned IC-RT-PCR products confirmed they were part of the CsCMV replicase gene, indicating that the virus was still present after thermotherapy and meristem-tip culture. PTA-ELISA enabled initial screening of virus-positive cassava, reducing the number of plants to be further analyzed by IC-RT-PCR. Though CsCMV has been considered of minor importance, its widespread nature, as noticed in Paraná, indicates the need for adoption of effective control measures.Sociedade Brasileira de Fitopatologia2011-10-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S1982-56762011000500001Tropical Plant Pathology v.36 n.5 2011reponame:Tropical plant pathology (Online)instname:Sociedade Brasileira de Fitopatologiainstacron:SBF10.1590/S1982-56762011000500001info:eu-repo/semantics/openAccessSilva,Jaqueline M.Carnelossi,Patrícia R.Bijora,TaiseFacco,Cassiele U.Picoli,Marcelo H.S.Souto,Eliezer R.Oliveira,Arildo J.B.Almeida,Álvaro M.R.eng2012-01-31T00:00:00Zoai:scielo:S1982-56762011000500001Revistahttps://www.scielo.br/j/tpp/ONGhttps://old.scielo.br/oai/scielo-oai.phpsbf-revista@ufla.br1983-20521982-5676opendoar:2012-01-31T00:00Tropical plant pathology (Online) - Sociedade Brasileira de Fitopatologiafalse
dc.title.none.fl_str_mv Immunocapture-RT-PCR detection of Cassava common mosaic virus in cassava obtained from meristem-tip culture in Paraná state
title Immunocapture-RT-PCR detection of Cassava common mosaic virus in cassava obtained from meristem-tip culture in Paraná state
spellingShingle Immunocapture-RT-PCR detection of Cassava common mosaic virus in cassava obtained from meristem-tip culture in Paraná state
Silva,Jaqueline M.
Potexvirus
Euphorbiaceae
IC-RT-PCR
indexing
PTA-ELISA
title_short Immunocapture-RT-PCR detection of Cassava common mosaic virus in cassava obtained from meristem-tip culture in Paraná state
title_full Immunocapture-RT-PCR detection of Cassava common mosaic virus in cassava obtained from meristem-tip culture in Paraná state
title_fullStr Immunocapture-RT-PCR detection of Cassava common mosaic virus in cassava obtained from meristem-tip culture in Paraná state
title_full_unstemmed Immunocapture-RT-PCR detection of Cassava common mosaic virus in cassava obtained from meristem-tip culture in Paraná state
title_sort Immunocapture-RT-PCR detection of Cassava common mosaic virus in cassava obtained from meristem-tip culture in Paraná state
author Silva,Jaqueline M.
author_facet Silva,Jaqueline M.
Carnelossi,Patrícia R.
Bijora,Taise
Facco,Cassiele U.
Picoli,Marcelo H.S.
Souto,Eliezer R.
Oliveira,Arildo J.B.
Almeida,Álvaro M.R.
author_role author
author2 Carnelossi,Patrícia R.
Bijora,Taise
Facco,Cassiele U.
Picoli,Marcelo H.S.
Souto,Eliezer R.
Oliveira,Arildo J.B.
Almeida,Álvaro M.R.
author2_role author
author
author
author
author
author
author
dc.contributor.author.fl_str_mv Silva,Jaqueline M.
Carnelossi,Patrícia R.
Bijora,Taise
Facco,Cassiele U.
Picoli,Marcelo H.S.
Souto,Eliezer R.
Oliveira,Arildo J.B.
Almeida,Álvaro M.R.
dc.subject.por.fl_str_mv Potexvirus
Euphorbiaceae
IC-RT-PCR
indexing
PTA-ELISA
topic Potexvirus
Euphorbiaceae
IC-RT-PCR
indexing
PTA-ELISA
description A virus survey conducted in the northwest region of Paraná, the main cassava-producing region of that state, showed Cassava common mosaic virus (CsCMV) to be widespread, infecting more than 90% of plants from all cassava cultivars. A CsCMV isolate was purified and used to raise a high-titer (1/1.000) polyclonal antiserum for indexing plants produced from meristem-tip culture, using PTA-ELISA. From an initial production of 110 plants of cultivar Olho Junto, 31 remained infected as indicated by PTA-ELISA. To improve the sensitivity of virus detection, an immunocapture-RT-PCR (IC-RT-PCR) protocol was established. Virus-specific IgG, purified and combined with a primer set for the genus Potexvirus, could readily detect CsCMV in cassava crude leaf extracts. IC-RT-PCR products of 750 bp were amplified from six out of 35 plants previously tested as virus-negative by PTA-ELISA. Sequence analysis of cloned IC-RT-PCR products confirmed they were part of the CsCMV replicase gene, indicating that the virus was still present after thermotherapy and meristem-tip culture. PTA-ELISA enabled initial screening of virus-positive cassava, reducing the number of plants to be further analyzed by IC-RT-PCR. Though CsCMV has been considered of minor importance, its widespread nature, as noticed in Paraná, indicates the need for adoption of effective control measures.
publishDate 2011
dc.date.none.fl_str_mv 2011-10-01
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1982-56762011000500001
url http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1982-56762011000500001
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv 10.1590/S1982-56762011000500001
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv text/html
dc.publisher.none.fl_str_mv Sociedade Brasileira de Fitopatologia
publisher.none.fl_str_mv Sociedade Brasileira de Fitopatologia
dc.source.none.fl_str_mv Tropical Plant Pathology v.36 n.5 2011
reponame:Tropical plant pathology (Online)
instname:Sociedade Brasileira de Fitopatologia
instacron:SBF
instname_str Sociedade Brasileira de Fitopatologia
instacron_str SBF
institution SBF
reponame_str Tropical plant pathology (Online)
collection Tropical plant pathology (Online)
repository.name.fl_str_mv Tropical plant pathology (Online) - Sociedade Brasileira de Fitopatologia
repository.mail.fl_str_mv sbf-revista@ufla.br
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