Development of a scar marker for Pierce's Disease strains of Xylella fastidiosa
Autor(a) principal: | |
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Data de Publicação: | 2005 |
Outros Autores: | , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Fitopatologia Brasileira |
Texto Completo: | http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0100-41582005000200002 |
Resumo: | The objective of this research was to develop a primer for a polymerase chain reaction specific for Xylella fastidiosa strains that cause Pierce's Disease (PD) in grapes (Vitis vinifera). The DNA amplification of 23 different strains of X. fastidiosa, using a set of primers REP1-R (5'-IIIICGICGIATCCIGGC-3') and REP 2 (5'-ICGICTTATCIGGCCTAC-3') using the following program: 94 ºC/2 min; 35 X (94 ºC/1 min, 45 ºC/1 min and 72 ºC/1 min and 30 s) 72 ºC/5 min, produced a fragment of 630 bp that differentiated the strains that cause disease in grapes from the other strains. However, REP banding patterns could not be considered reliable for detection because the REP1-R and REP 2 primers correspond to repetitive sequences, which are found throughout the bacterial genome. The amplified product of 630 bp was eluted from the agarose gel, purified and sequenced. The nucleotide sequence information was used to identify and synthesize an specific oligonucleotide for X. fastidiosa strains that cause Pierce's Disease denominated Xf-1 (5'-CGGGGGTGTAGGAGGGGTTGT-3') which was used jointly with the REP-2 primer at the following conditions: 94 ºC/2 min; 35 X (94 ºC/1 min, 62 ºC/1 min; 72 ºC/1 min and 30 s) 72 ºC/10 min. The DNAs isolated from strains of X. fastidiosa from other hosts [almond (Prumus amygdalus), citrus (Citrus spp.), coffee (Coffea arabica), elm (Ulmus americana), mulberry (Morus rubra), oak (Quercus rubra), periwinkle wilt (Catharantus roseus), plums (Prunus salicina) and ragweed (Ambrosia artemisiifolia)] and also from other Gram negative and positive bacteria were submitted to amplification with a pair of primers Xf-1/REP 2 to verify its specificity. A fragment, about 350 bp, was amplified only when the DNA from strains of X. fastidiosa isolated from grapes was employed. |
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Development of a scar marker for Pierce's Disease strains of Xylella fastidiosaoligonucleotidepolymerase chain reactionPCRREP-PCRThe objective of this research was to develop a primer for a polymerase chain reaction specific for Xylella fastidiosa strains that cause Pierce's Disease (PD) in grapes (Vitis vinifera). The DNA amplification of 23 different strains of X. fastidiosa, using a set of primers REP1-R (5'-IIIICGICGIATCCIGGC-3') and REP 2 (5'-ICGICTTATCIGGCCTAC-3') using the following program: 94 ºC/2 min; 35 X (94 ºC/1 min, 45 ºC/1 min and 72 ºC/1 min and 30 s) 72 ºC/5 min, produced a fragment of 630 bp that differentiated the strains that cause disease in grapes from the other strains. However, REP banding patterns could not be considered reliable for detection because the REP1-R and REP 2 primers correspond to repetitive sequences, which are found throughout the bacterial genome. The amplified product of 630 bp was eluted from the agarose gel, purified and sequenced. The nucleotide sequence information was used to identify and synthesize an specific oligonucleotide for X. fastidiosa strains that cause Pierce's Disease denominated Xf-1 (5'-CGGGGGTGTAGGAGGGGTTGT-3') which was used jointly with the REP-2 primer at the following conditions: 94 ºC/2 min; 35 X (94 ºC/1 min, 62 ºC/1 min; 72 ºC/1 min and 30 s) 72 ºC/10 min. The DNAs isolated from strains of X. fastidiosa from other hosts [almond (Prumus amygdalus), citrus (Citrus spp.), coffee (Coffea arabica), elm (Ulmus americana), mulberry (Morus rubra), oak (Quercus rubra), periwinkle wilt (Catharantus roseus), plums (Prunus salicina) and ragweed (Ambrosia artemisiifolia)] and also from other Gram negative and positive bacteria were submitted to amplification with a pair of primers Xf-1/REP 2 to verify its specificity. A fragment, about 350 bp, was amplified only when the DNA from strains of X. fastidiosa isolated from grapes was employed.Sociedade Brasileira de Fitopatologia2005-04-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S0100-41582005000200002Fitopatologia Brasileira v.30 n.2 2005reponame:Fitopatologia Brasileirainstname:Sociedade Brasileira de Fitopatologia (SBF)instacron:SBF10.1590/S0100-41582005000200002info:eu-repo/semantics/openAccessTravensolo,Regiane F.Ciapina,Luciane P.Lemos,Eliana G. M.eng2005-05-16T00:00:00Zoai:scielo:S0100-41582005000200002Revistahttp://www.scielo.br/fbONGhttps://old.scielo.br/oai/scielo-oai.php||sbf-revista@ufla.br1678-46770100-4158opendoar:2005-05-16T00:00Fitopatologia Brasileira - Sociedade Brasileira de Fitopatologia (SBF)false |
dc.title.none.fl_str_mv |
Development of a scar marker for Pierce's Disease strains of Xylella fastidiosa |
title |
Development of a scar marker for Pierce's Disease strains of Xylella fastidiosa |
spellingShingle |
Development of a scar marker for Pierce's Disease strains of Xylella fastidiosa Travensolo,Regiane F. oligonucleotide polymerase chain reaction PCR REP-PCR |
title_short |
Development of a scar marker for Pierce's Disease strains of Xylella fastidiosa |
title_full |
Development of a scar marker for Pierce's Disease strains of Xylella fastidiosa |
title_fullStr |
Development of a scar marker for Pierce's Disease strains of Xylella fastidiosa |
title_full_unstemmed |
Development of a scar marker for Pierce's Disease strains of Xylella fastidiosa |
title_sort |
Development of a scar marker for Pierce's Disease strains of Xylella fastidiosa |
author |
Travensolo,Regiane F. |
author_facet |
Travensolo,Regiane F. Ciapina,Luciane P. Lemos,Eliana G. M. |
author_role |
author |
author2 |
Ciapina,Luciane P. Lemos,Eliana G. M. |
author2_role |
author author |
dc.contributor.author.fl_str_mv |
Travensolo,Regiane F. Ciapina,Luciane P. Lemos,Eliana G. M. |
dc.subject.por.fl_str_mv |
oligonucleotide polymerase chain reaction PCR REP-PCR |
topic |
oligonucleotide polymerase chain reaction PCR REP-PCR |
description |
The objective of this research was to develop a primer for a polymerase chain reaction specific for Xylella fastidiosa strains that cause Pierce's Disease (PD) in grapes (Vitis vinifera). The DNA amplification of 23 different strains of X. fastidiosa, using a set of primers REP1-R (5'-IIIICGICGIATCCIGGC-3') and REP 2 (5'-ICGICTTATCIGGCCTAC-3') using the following program: 94 ºC/2 min; 35 X (94 ºC/1 min, 45 ºC/1 min and 72 ºC/1 min and 30 s) 72 ºC/5 min, produced a fragment of 630 bp that differentiated the strains that cause disease in grapes from the other strains. However, REP banding patterns could not be considered reliable for detection because the REP1-R and REP 2 primers correspond to repetitive sequences, which are found throughout the bacterial genome. The amplified product of 630 bp was eluted from the agarose gel, purified and sequenced. The nucleotide sequence information was used to identify and synthesize an specific oligonucleotide for X. fastidiosa strains that cause Pierce's Disease denominated Xf-1 (5'-CGGGGGTGTAGGAGGGGTTGT-3') which was used jointly with the REP-2 primer at the following conditions: 94 ºC/2 min; 35 X (94 ºC/1 min, 62 ºC/1 min; 72 ºC/1 min and 30 s) 72 ºC/10 min. The DNAs isolated from strains of X. fastidiosa from other hosts [almond (Prumus amygdalus), citrus (Citrus spp.), coffee (Coffea arabica), elm (Ulmus americana), mulberry (Morus rubra), oak (Quercus rubra), periwinkle wilt (Catharantus roseus), plums (Prunus salicina) and ragweed (Ambrosia artemisiifolia)] and also from other Gram negative and positive bacteria were submitted to amplification with a pair of primers Xf-1/REP 2 to verify its specificity. A fragment, about 350 bp, was amplified only when the DNA from strains of X. fastidiosa isolated from grapes was employed. |
publishDate |
2005 |
dc.date.none.fl_str_mv |
2005-04-01 |
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=S0100-41582005000200002 |
url |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0100-41582005000200002 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
10.1590/S0100-41582005000200002 |
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 |
Fitopatologia Brasileira v.30 n.2 2005 reponame:Fitopatologia Brasileira instname:Sociedade Brasileira de Fitopatologia (SBF) instacron:SBF |
instname_str |
Sociedade Brasileira de Fitopatologia (SBF) |
instacron_str |
SBF |
institution |
SBF |
reponame_str |
Fitopatologia Brasileira |
collection |
Fitopatologia Brasileira |
repository.name.fl_str_mv |
Fitopatologia Brasileira - Sociedade Brasileira de Fitopatologia (SBF) |
repository.mail.fl_str_mv |
||sbf-revista@ufla.br |
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