Protocol for detecting Curtobacterium flaccumfaciens pv. flaccumfaciens in the leaves of infected bean plants (Phaseolus vulgaris L.)
Autor(a) principal: | |
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Data de Publicação: | 2021 |
Outros Autores: | , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Acta Scientiarum. Agronomy (Online) |
Texto Completo: | http://www.periodicos.uem.br/ojs/index.php/ActaSciAgron/article/view/51031 |
Resumo: | Curtobacterium flaccumfaciens pv. flaccumfaciens (Cff), which is a gram-positive bacterium that causes wilting in several cultivated plants, is one of the main disease-causing bacteria in beans (Phaseolus vulgaris L.). Different techniques are employed to detect pathogens in bean plants; however, only a few slow and inefficient methodologies can be used to detect Cff. Hence, there is a need for rapid and efficient techniques to detect Cff for disease management. The objective of our study was to develop a technique to effectively detect Cff in the leaves of infected bean plants by optimizing a previously developed method. We modified the extraction method by reducing the amount of water used, replacing seeds with leaf tissues (obtained from the veins and petioles) and using CffFOR2 and CffREV4 primers specific for Cff. Our optimized method exploits a concentration gradient generated in the plant tissue through osmosis to disrupt the plant cell wall and efficiently isolate bacterial cells. DNA extracted from the isolated bacterial cells was amplified by polymerase chain reaction. This method allowed efficient detection of Cff in leaf tissues. |
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Acta Scientiarum. Agronomy (Online) |
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Protocol for detecting Curtobacterium flaccumfaciens pv. flaccumfaciens in the leaves of infected bean plants (Phaseolus vulgaris L.)Protocol for detecting Curtobacterium flaccumfaciens pv. flaccumfaciens in the leaves of infected bean plants (Phaseolus vulgaris L.)Curtobacterium wilt; DNA extraction; leaf tissue; bacterium; common beans.Curtobacterium wilt; DNA extraction; leaf tissue; bacterium; common beans.Curtobacterium flaccumfaciens pv. flaccumfaciens (Cff), which is a gram-positive bacterium that causes wilting in several cultivated plants, is one of the main disease-causing bacteria in beans (Phaseolus vulgaris L.). Different techniques are employed to detect pathogens in bean plants; however, only a few slow and inefficient methodologies can be used to detect Cff. Hence, there is a need for rapid and efficient techniques to detect Cff for disease management. The objective of our study was to develop a technique to effectively detect Cff in the leaves of infected bean plants by optimizing a previously developed method. We modified the extraction method by reducing the amount of water used, replacing seeds with leaf tissues (obtained from the veins and petioles) and using CffFOR2 and CffREV4 primers specific for Cff. Our optimized method exploits a concentration gradient generated in the plant tissue through osmosis to disrupt the plant cell wall and efficiently isolate bacterial cells. DNA extracted from the isolated bacterial cells was amplified by polymerase chain reaction. This method allowed efficient detection of Cff in leaf tissues.Curtobacterium flaccumfaciens pv. flaccumfaciens (Cff), which is a gram-positive bacterium that causes wilting in several cultivated plants, is one of the main disease-causing bacteria in beans (Phaseolus vulgaris L.). Different techniques are employed to detect pathogens in bean plants; however, only a few slow and inefficient methodologies can be used to detect Cff. Hence, there is a need for rapid and efficient techniques to detect Cff for disease management. The objective of our study was to develop a technique to effectively detect Cff in the leaves of infected bean plants by optimizing a previously developed method. We modified the extraction method by reducing the amount of water used, replacing seeds with leaf tissues (obtained from the veins and petioles) and using CffFOR2 and CffREV4 primers specific for Cff. Our optimized method exploits a concentration gradient generated in the plant tissue through osmosis to disrupt the plant cell wall and efficiently isolate bacterial cells. DNA extracted from the isolated bacterial cells was amplified by polymerase chain reaction. This method allowed efficient detection of Cff in leaf tissues.Universidade Estadual de Maringá2021-03-18info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfhttp://www.periodicos.uem.br/ojs/index.php/ActaSciAgron/article/view/5103110.4025/actasciagron.v43i1.51031Acta Scientiarum. Agronomy; Vol 43 (2021): Publicação contínua; e51031Acta Scientiarum. Agronomy; v. 43 (2021): Publicação contínua; e510311807-86211679-9275reponame:Acta Scientiarum. Agronomy (Online)instname:Universidade Estadual de Maringá (UEM)instacron:UEMenghttp://www.periodicos.uem.br/ojs/index.php/ActaSciAgron/article/view/51031/751375151805Copyright (c) 2021 Acta Scientiarum. Agronomyhttps://creativecommons.org/licenses/by-nc-nd/4.0info:eu-repo/semantics/openAccessPuia, Jacqueline Dalbelo Murari, Rafaela Rodrigues Borsato, Leandro Camargo Sugahara, Vanessa Hitomi Machineski, Gabriela da Silva Canteri, Marcelo Giovanetti Vigo, Sandra Cristina 2021-07-27T17:52:14Zoai:periodicos.uem.br/ojs:article/51031Revistahttp://www.periodicos.uem.br/ojs/index.php/ActaSciAgronPUBhttp://www.periodicos.uem.br/ojs/index.php/ActaSciAgron/oaiactaagron@uem.br||actaagron@uem.br|| edamasio@uem.br1807-86211679-9275opendoar:2021-07-27T17:52:14Acta Scientiarum. Agronomy (Online) - Universidade Estadual de Maringá (UEM)false |
dc.title.none.fl_str_mv |
Protocol for detecting Curtobacterium flaccumfaciens pv. flaccumfaciens in the leaves of infected bean plants (Phaseolus vulgaris L.) Protocol for detecting Curtobacterium flaccumfaciens pv. flaccumfaciens in the leaves of infected bean plants (Phaseolus vulgaris L.) |
title |
Protocol for detecting Curtobacterium flaccumfaciens pv. flaccumfaciens in the leaves of infected bean plants (Phaseolus vulgaris L.) |
spellingShingle |
Protocol for detecting Curtobacterium flaccumfaciens pv. flaccumfaciens in the leaves of infected bean plants (Phaseolus vulgaris L.) Puia, Jacqueline Dalbelo Curtobacterium wilt; DNA extraction; leaf tissue; bacterium; common beans. Curtobacterium wilt; DNA extraction; leaf tissue; bacterium; common beans. |
title_short |
Protocol for detecting Curtobacterium flaccumfaciens pv. flaccumfaciens in the leaves of infected bean plants (Phaseolus vulgaris L.) |
title_full |
Protocol for detecting Curtobacterium flaccumfaciens pv. flaccumfaciens in the leaves of infected bean plants (Phaseolus vulgaris L.) |
title_fullStr |
Protocol for detecting Curtobacterium flaccumfaciens pv. flaccumfaciens in the leaves of infected bean plants (Phaseolus vulgaris L.) |
title_full_unstemmed |
Protocol for detecting Curtobacterium flaccumfaciens pv. flaccumfaciens in the leaves of infected bean plants (Phaseolus vulgaris L.) |
title_sort |
Protocol for detecting Curtobacterium flaccumfaciens pv. flaccumfaciens in the leaves of infected bean plants (Phaseolus vulgaris L.) |
author |
Puia, Jacqueline Dalbelo |
author_facet |
Puia, Jacqueline Dalbelo Murari, Rafaela Rodrigues Borsato, Leandro Camargo Sugahara, Vanessa Hitomi Machineski, Gabriela da Silva Canteri, Marcelo Giovanetti Vigo, Sandra Cristina |
author_role |
author |
author2 |
Murari, Rafaela Rodrigues Borsato, Leandro Camargo Sugahara, Vanessa Hitomi Machineski, Gabriela da Silva Canteri, Marcelo Giovanetti Vigo, Sandra Cristina |
author2_role |
author author author author author author |
dc.contributor.author.fl_str_mv |
Puia, Jacqueline Dalbelo Murari, Rafaela Rodrigues Borsato, Leandro Camargo Sugahara, Vanessa Hitomi Machineski, Gabriela da Silva Canteri, Marcelo Giovanetti Vigo, Sandra Cristina |
dc.subject.por.fl_str_mv |
Curtobacterium wilt; DNA extraction; leaf tissue; bacterium; common beans. Curtobacterium wilt; DNA extraction; leaf tissue; bacterium; common beans. |
topic |
Curtobacterium wilt; DNA extraction; leaf tissue; bacterium; common beans. Curtobacterium wilt; DNA extraction; leaf tissue; bacterium; common beans. |
description |
Curtobacterium flaccumfaciens pv. flaccumfaciens (Cff), which is a gram-positive bacterium that causes wilting in several cultivated plants, is one of the main disease-causing bacteria in beans (Phaseolus vulgaris L.). Different techniques are employed to detect pathogens in bean plants; however, only a few slow and inefficient methodologies can be used to detect Cff. Hence, there is a need for rapid and efficient techniques to detect Cff for disease management. The objective of our study was to develop a technique to effectively detect Cff in the leaves of infected bean plants by optimizing a previously developed method. We modified the extraction method by reducing the amount of water used, replacing seeds with leaf tissues (obtained from the veins and petioles) and using CffFOR2 and CffREV4 primers specific for Cff. Our optimized method exploits a concentration gradient generated in the plant tissue through osmosis to disrupt the plant cell wall and efficiently isolate bacterial cells. DNA extracted from the isolated bacterial cells was amplified by polymerase chain reaction. This method allowed efficient detection of Cff in leaf tissues. |
publishDate |
2021 |
dc.date.none.fl_str_mv |
2021-03-18 |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://www.periodicos.uem.br/ojs/index.php/ActaSciAgron/article/view/51031 10.4025/actasciagron.v43i1.51031 |
url |
http://www.periodicos.uem.br/ojs/index.php/ActaSciAgron/article/view/51031 |
identifier_str_mv |
10.4025/actasciagron.v43i1.51031 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
http://www.periodicos.uem.br/ojs/index.php/ActaSciAgron/article/view/51031/751375151805 |
dc.rights.driver.fl_str_mv |
Copyright (c) 2021 Acta Scientiarum. Agronomy https://creativecommons.org/licenses/by-nc-nd/4.0 info:eu-repo/semantics/openAccess |
rights_invalid_str_mv |
Copyright (c) 2021 Acta Scientiarum. Agronomy https://creativecommons.org/licenses/by-nc-nd/4.0 |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Universidade Estadual de Maringá |
publisher.none.fl_str_mv |
Universidade Estadual de Maringá |
dc.source.none.fl_str_mv |
Acta Scientiarum. Agronomy; Vol 43 (2021): Publicação contínua; e51031 Acta Scientiarum. Agronomy; v. 43 (2021): Publicação contínua; e51031 1807-8621 1679-9275 reponame:Acta Scientiarum. Agronomy (Online) instname:Universidade Estadual de Maringá (UEM) instacron:UEM |
instname_str |
Universidade Estadual de Maringá (UEM) |
instacron_str |
UEM |
institution |
UEM |
reponame_str |
Acta Scientiarum. Agronomy (Online) |
collection |
Acta Scientiarum. Agronomy (Online) |
repository.name.fl_str_mv |
Acta Scientiarum. Agronomy (Online) - Universidade Estadual de Maringá (UEM) |
repository.mail.fl_str_mv |
actaagron@uem.br||actaagron@uem.br|| edamasio@uem.br |
_version_ |
1799305911413506048 |