Immunocapture-RT-PCR detection of Cassava common mosaic virus in cassava obtained from meristem-tip culture in Paraná state
Autor(a) principal: | |
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Data de Publicação: | 2011 |
Outros Autores: | , , , , , , |
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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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 |
_version_ |
1754824585533980672 |