Detection of Xanthomonas axonopodis pv. phaseoli in bean seeds by flow cytometry, immunostaining and direct viable counting
Autor(a) principal: | |
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Data de Publicação: | 2010 |
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=S1982-56762010000400002 |
Resumo: | Flow cytometric analysis of immuno-stained cells (immuno-FCM) was compared to immunofluorescence microscopy (IF) and dilution plating on a semi-selective medium, for quantitative detection of Xanthomonas axonopodis pv. phaseoli (Xap) in bean seed extracts. Cell concentrations of Xap between 10³-10(7) CFU/mL were added to healthy bean seed extracts. A flow cytometry sorting procedure was developed to separate immuno-stained Xap cells from crude seed extracts and confirming by PCR. FCM was evaluated for direct viable counting (DVC) of Xap using combinations of propidium iodide (PI) and carboxy fluorescein diacetate (cFDA) or PI and SYTO 9 and also the combination of immuno-FCM and PI. Dilution plating and IF allowed detection of Xap in bean seed extracts in a range of 10³-10(6) CFU/mL and immuno-FCM from 10(4)-10(6) CFU/mL. Sorted cells could be detected in crude seed extracts by PCR without further extraction. FCM also allowed quantification of viable cells of Xap after DVC procedures; the red fluorescent dye propidium iodide was used to identify dead cells in combination with the green fluorescent dyes cFDA or SYTO 9, these identifying live cells. The combination of immuno-FCM and PI could be more promising and reliable to detect this pathogen in seeds. |
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Tropical plant pathology (Online) |
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Detection of Xanthomonas axonopodis pv. phaseoli in bean seeds by flow cytometry, immunostaining and direct viable countingseed pathologyflow sortingPCR-amplificationviability probesimmunofluorescencebacteriaFlow cytometric analysis of immuno-stained cells (immuno-FCM) was compared to immunofluorescence microscopy (IF) and dilution plating on a semi-selective medium, for quantitative detection of Xanthomonas axonopodis pv. phaseoli (Xap) in bean seed extracts. Cell concentrations of Xap between 10³-10(7) CFU/mL were added to healthy bean seed extracts. A flow cytometry sorting procedure was developed to separate immuno-stained Xap cells from crude seed extracts and confirming by PCR. FCM was evaluated for direct viable counting (DVC) of Xap using combinations of propidium iodide (PI) and carboxy fluorescein diacetate (cFDA) or PI and SYTO 9 and also the combination of immuno-FCM and PI. Dilution plating and IF allowed detection of Xap in bean seed extracts in a range of 10³-10(6) CFU/mL and immuno-FCM from 10(4)-10(6) CFU/mL. Sorted cells could be detected in crude seed extracts by PCR without further extraction. FCM also allowed quantification of viable cells of Xap after DVC procedures; the red fluorescent dye propidium iodide was used to identify dead cells in combination with the green fluorescent dyes cFDA or SYTO 9, these identifying live cells. The combination of immuno-FCM and PI could be more promising and reliable to detect this pathogen in seeds.Sociedade Brasileira de Fitopatologia2010-08-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S1982-56762010000400002Tropical Plant Pathology v.35 n.4 2010reponame:Tropical plant pathology (Online)instname:Sociedade Brasileira de Fitopatologiainstacron:SBF10.1590/S1982-56762010000400002info:eu-repo/semantics/openAccessTebaldi,Nilvanira DPeters,JeroenSouza,Ricardo MChitarra,Luiz Gvan der Zouwen,PatriciaBergervoet,Janvan der Wolf,Janeng2010-11-03T00:00:00Zoai:scielo:S1982-56762010000400002Revistahttps://www.scielo.br/j/tpp/ONGhttps://old.scielo.br/oai/scielo-oai.phpsbf-revista@ufla.br1983-20521982-5676opendoar:2010-11-03T00:00Tropical plant pathology (Online) - Sociedade Brasileira de Fitopatologiafalse |
dc.title.none.fl_str_mv |
Detection of Xanthomonas axonopodis pv. phaseoli in bean seeds by flow cytometry, immunostaining and direct viable counting |
title |
Detection of Xanthomonas axonopodis pv. phaseoli in bean seeds by flow cytometry, immunostaining and direct viable counting |
spellingShingle |
Detection of Xanthomonas axonopodis pv. phaseoli in bean seeds by flow cytometry, immunostaining and direct viable counting Tebaldi,Nilvanira D seed pathology flow sorting PCR-amplification viability probes immunofluorescence bacteria |
title_short |
Detection of Xanthomonas axonopodis pv. phaseoli in bean seeds by flow cytometry, immunostaining and direct viable counting |
title_full |
Detection of Xanthomonas axonopodis pv. phaseoli in bean seeds by flow cytometry, immunostaining and direct viable counting |
title_fullStr |
Detection of Xanthomonas axonopodis pv. phaseoli in bean seeds by flow cytometry, immunostaining and direct viable counting |
title_full_unstemmed |
Detection of Xanthomonas axonopodis pv. phaseoli in bean seeds by flow cytometry, immunostaining and direct viable counting |
title_sort |
Detection of Xanthomonas axonopodis pv. phaseoli in bean seeds by flow cytometry, immunostaining and direct viable counting |
author |
Tebaldi,Nilvanira D |
author_facet |
Tebaldi,Nilvanira D Peters,Jeroen Souza,Ricardo M Chitarra,Luiz G van der Zouwen,Patricia Bergervoet,Jan van der Wolf,Jan |
author_role |
author |
author2 |
Peters,Jeroen Souza,Ricardo M Chitarra,Luiz G van der Zouwen,Patricia Bergervoet,Jan van der Wolf,Jan |
author2_role |
author author author author author author |
dc.contributor.author.fl_str_mv |
Tebaldi,Nilvanira D Peters,Jeroen Souza,Ricardo M Chitarra,Luiz G van der Zouwen,Patricia Bergervoet,Jan van der Wolf,Jan |
dc.subject.por.fl_str_mv |
seed pathology flow sorting PCR-amplification viability probes immunofluorescence bacteria |
topic |
seed pathology flow sorting PCR-amplification viability probes immunofluorescence bacteria |
description |
Flow cytometric analysis of immuno-stained cells (immuno-FCM) was compared to immunofluorescence microscopy (IF) and dilution plating on a semi-selective medium, for quantitative detection of Xanthomonas axonopodis pv. phaseoli (Xap) in bean seed extracts. Cell concentrations of Xap between 10³-10(7) CFU/mL were added to healthy bean seed extracts. A flow cytometry sorting procedure was developed to separate immuno-stained Xap cells from crude seed extracts and confirming by PCR. FCM was evaluated for direct viable counting (DVC) of Xap using combinations of propidium iodide (PI) and carboxy fluorescein diacetate (cFDA) or PI and SYTO 9 and also the combination of immuno-FCM and PI. Dilution plating and IF allowed detection of Xap in bean seed extracts in a range of 10³-10(6) CFU/mL and immuno-FCM from 10(4)-10(6) CFU/mL. Sorted cells could be detected in crude seed extracts by PCR without further extraction. FCM also allowed quantification of viable cells of Xap after DVC procedures; the red fluorescent dye propidium iodide was used to identify dead cells in combination with the green fluorescent dyes cFDA or SYTO 9, these identifying live cells. The combination of immuno-FCM and PI could be more promising and reliable to detect this pathogen in seeds. |
publishDate |
2010 |
dc.date.none.fl_str_mv |
2010-08-01 |
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=S1982-56762010000400002 |
url |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1982-56762010000400002 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
10.1590/S1982-56762010000400002 |
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.35 n.4 2010 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_ |
1754824585354674176 |