Real-time PCR for detection and quantification of Xanthomonas axonopodis pv. dieffenbachiae on Anthurium
Effective control of Xanthomonas axonopodis pv. dieffenbachiae, the causal agent of anthurium bacterial blight, requires efficient screening of quarantine plants against this pathogen. A reproducible real-time PCR method that targets a portion of a gene from the LPS cluster, encoding a putative ABC transporter, was developed to detect X. axonopodis pv. dieffenbachiae in anthurium samples. The specificity of the assay was confirmed with a panel of strains belonging to X. axonopodis pv dieffenbachiae and other pathovars and species. The assay was conducted with serial 10-fold dilutions of purified bacterial DNA and pure cultures (12 strains tested). It was possible to construct standard curves with a high correlation coefficient (r2 = 0.99) in the range of 15 ng to 1.5 pg for purified DNA and 107 to 102 CFU/ml for bacterial cultures. DNA extraction methods were compared for maximum DNA yield from anthurium plants. Homogenization of tissues with a Homex grinder (Bioreba) followed by purification using the DNeasy blood and tissue kit (Qiagen) was the most efficient method. The assay was successfully used to monitor the in vivo infection of anthurium experimentally inoculated with X. axonopodis pv. dieffenbachiae. This Real-Time PCR assay provides a rapid method for the early detection of the bacterium in symptomless anthurium tissues. It should therefore be a very useful diagnostic tool for indexing propagation material in nurseries and for surveillance of international movement of X. axonopodis pv. dieffenbachiae on anthurium. (Texte intégral)
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Subjects: | H20 - Maladies des plantes, Xanthomonas, PCR, analyse quantitative, identification, Anthurium, diagnostic précoce, contrôle de maladies, http://aims.fao.org/aos/agrovoc/c_8455, http://aims.fao.org/aos/agrovoc/c_34079, http://aims.fao.org/aos/agrovoc/c_32660, http://aims.fao.org/aos/agrovoc/c_3791, http://aims.fao.org/aos/agrovoc/c_487, http://aims.fao.org/aos/agrovoc/c_36584, http://aims.fao.org/aos/agrovoc/c_2327, http://aims.fao.org/aos/agrovoc/c_6543, http://aims.fao.org/aos/agrovoc/c_3081, |
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dig-cirad-fr-5472042024-01-28T16:33:56Z http://agritrop.cirad.fr/547204/ http://agritrop.cirad.fr/547204/ Real-time PCR for detection and quantification of Xanthomonas axonopodis pv. dieffenbachiae on Anthurium. Péréfarres Frédéric, Jouen Emmanuel, Soustrade Isabelle, Gagnevin Lionel, Laurent Annie, Pruvost Olivier. 2008. Journal of Plant Pathology, 90 (2), suppl., Résumé, S2.306 International Congress of Plant Pathology. 9, Turin, Italie, 24 Août 2008/29 Août 2008.http://www.sipav.org/main/jpp/volumes/0808/ICPP_2008.pdf <http://www.sipav.org/main/jpp/volumes/0808/ICPP_2008.pdf> Real-time PCR for detection and quantification of Xanthomonas axonopodis pv. dieffenbachiae on Anthurium Péréfarres, Frédéric Jouen, Emmanuel Soustrade, Isabelle Gagnevin, Lionel Laurent, Annie Pruvost, Olivier eng 2008 Journal of Plant Pathology H20 - Maladies des plantes Xanthomonas PCR analyse quantitative identification Anthurium diagnostic précoce contrôle de maladies http://aims.fao.org/aos/agrovoc/c_8455 http://aims.fao.org/aos/agrovoc/c_34079 http://aims.fao.org/aos/agrovoc/c_32660 http://aims.fao.org/aos/agrovoc/c_3791 http://aims.fao.org/aos/agrovoc/c_487 http://aims.fao.org/aos/agrovoc/c_36584 http://aims.fao.org/aos/agrovoc/c_2327 La Réunion France http://aims.fao.org/aos/agrovoc/c_6543 http://aims.fao.org/aos/agrovoc/c_3081 Effective control of Xanthomonas axonopodis pv. dieffenbachiae, the causal agent of anthurium bacterial blight, requires efficient screening of quarantine plants against this pathogen. A reproducible real-time PCR method that targets a portion of a gene from the LPS cluster, encoding a putative ABC transporter, was developed to detect X. axonopodis pv. dieffenbachiae in anthurium samples. The specificity of the assay was confirmed with a panel of strains belonging to X. axonopodis pv dieffenbachiae and other pathovars and species. The assay was conducted with serial 10-fold dilutions of purified bacterial DNA and pure cultures (12 strains tested). It was possible to construct standard curves with a high correlation coefficient (r2 = 0.99) in the range of 15 ng to 1.5 pg for purified DNA and 107 to 102 CFU/ml for bacterial cultures. DNA extraction methods were compared for maximum DNA yield from anthurium plants. Homogenization of tissues with a Homex grinder (Bioreba) followed by purification using the DNeasy blood and tissue kit (Qiagen) was the most efficient method. The assay was successfully used to monitor the in vivo infection of anthurium experimentally inoculated with X. axonopodis pv. dieffenbachiae. This Real-Time PCR assay provides a rapid method for the early detection of the bacterium in symptomless anthurium tissues. It should therefore be a very useful diagnostic tool for indexing propagation material in nurseries and for surveillance of international movement of X. axonopodis pv. dieffenbachiae on anthurium. (Texte intégral) article info:eu-repo/semantics/article Journal Article info:eu-repo/semantics/closedAccess http://www.sipav.org/main/jpp/volumes/0808/ICPP_2008.pdf http://catalogue-bibliotheques.cirad.fr/cgi-bin/koha/opac-detail.pl?biblionumber=202222 info:eu-repo/semantics/altIdentifier/purl/http://www.sipav.org/main/jpp/volumes/0808/ICPP_2008.pdf |
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H20 - Maladies des plantes Xanthomonas PCR analyse quantitative identification Anthurium diagnostic précoce contrôle de maladies http://aims.fao.org/aos/agrovoc/c_8455 http://aims.fao.org/aos/agrovoc/c_34079 http://aims.fao.org/aos/agrovoc/c_32660 http://aims.fao.org/aos/agrovoc/c_3791 http://aims.fao.org/aos/agrovoc/c_487 http://aims.fao.org/aos/agrovoc/c_36584 http://aims.fao.org/aos/agrovoc/c_2327 http://aims.fao.org/aos/agrovoc/c_6543 http://aims.fao.org/aos/agrovoc/c_3081 H20 - Maladies des plantes Xanthomonas PCR analyse quantitative identification Anthurium diagnostic précoce contrôle de maladies http://aims.fao.org/aos/agrovoc/c_8455 http://aims.fao.org/aos/agrovoc/c_34079 http://aims.fao.org/aos/agrovoc/c_32660 http://aims.fao.org/aos/agrovoc/c_3791 http://aims.fao.org/aos/agrovoc/c_487 http://aims.fao.org/aos/agrovoc/c_36584 http://aims.fao.org/aos/agrovoc/c_2327 http://aims.fao.org/aos/agrovoc/c_6543 http://aims.fao.org/aos/agrovoc/c_3081 |
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H20 - Maladies des plantes Xanthomonas PCR analyse quantitative identification Anthurium diagnostic précoce contrôle de maladies http://aims.fao.org/aos/agrovoc/c_8455 http://aims.fao.org/aos/agrovoc/c_34079 http://aims.fao.org/aos/agrovoc/c_32660 http://aims.fao.org/aos/agrovoc/c_3791 http://aims.fao.org/aos/agrovoc/c_487 http://aims.fao.org/aos/agrovoc/c_36584 http://aims.fao.org/aos/agrovoc/c_2327 http://aims.fao.org/aos/agrovoc/c_6543 http://aims.fao.org/aos/agrovoc/c_3081 H20 - Maladies des plantes Xanthomonas PCR analyse quantitative identification Anthurium diagnostic précoce contrôle de maladies http://aims.fao.org/aos/agrovoc/c_8455 http://aims.fao.org/aos/agrovoc/c_34079 http://aims.fao.org/aos/agrovoc/c_32660 http://aims.fao.org/aos/agrovoc/c_3791 http://aims.fao.org/aos/agrovoc/c_487 http://aims.fao.org/aos/agrovoc/c_36584 http://aims.fao.org/aos/agrovoc/c_2327 http://aims.fao.org/aos/agrovoc/c_6543 http://aims.fao.org/aos/agrovoc/c_3081 Péréfarres, Frédéric Jouen, Emmanuel Soustrade, Isabelle Gagnevin, Lionel Laurent, Annie Pruvost, Olivier Real-time PCR for detection and quantification of Xanthomonas axonopodis pv. dieffenbachiae on Anthurium |
description |
Effective control of Xanthomonas axonopodis pv. dieffenbachiae, the causal agent of anthurium bacterial blight, requires efficient screening of quarantine plants against this pathogen. A reproducible real-time PCR method that targets a portion of a gene from the LPS cluster, encoding a putative ABC transporter, was developed to detect X. axonopodis pv. dieffenbachiae in anthurium samples. The specificity of the assay was confirmed with a panel of strains belonging to X. axonopodis pv dieffenbachiae and other pathovars and species. The assay was conducted with serial 10-fold dilutions of purified bacterial DNA and pure cultures (12 strains tested). It was possible to construct standard curves with a high correlation coefficient (r2 = 0.99) in the range of 15 ng to 1.5 pg for purified DNA and 107 to 102 CFU/ml for bacterial cultures. DNA extraction methods were compared for maximum DNA yield from anthurium plants. Homogenization of tissues with a Homex grinder (Bioreba) followed by purification using the DNeasy blood and tissue kit (Qiagen) was the most efficient method. The assay was successfully used to monitor the in vivo infection of anthurium experimentally inoculated with X. axonopodis pv. dieffenbachiae. This Real-Time PCR assay provides a rapid method for the early detection of the bacterium in symptomless anthurium tissues. It should therefore be a very useful diagnostic tool for indexing propagation material in nurseries and for surveillance of international movement of X. axonopodis pv. dieffenbachiae on anthurium. (Texte intégral) |
format |
article |
topic_facet |
H20 - Maladies des plantes Xanthomonas PCR analyse quantitative identification Anthurium diagnostic précoce contrôle de maladies http://aims.fao.org/aos/agrovoc/c_8455 http://aims.fao.org/aos/agrovoc/c_34079 http://aims.fao.org/aos/agrovoc/c_32660 http://aims.fao.org/aos/agrovoc/c_3791 http://aims.fao.org/aos/agrovoc/c_487 http://aims.fao.org/aos/agrovoc/c_36584 http://aims.fao.org/aos/agrovoc/c_2327 http://aims.fao.org/aos/agrovoc/c_6543 http://aims.fao.org/aos/agrovoc/c_3081 |
author |
Péréfarres, Frédéric Jouen, Emmanuel Soustrade, Isabelle Gagnevin, Lionel Laurent, Annie Pruvost, Olivier |
author_facet |
Péréfarres, Frédéric Jouen, Emmanuel Soustrade, Isabelle Gagnevin, Lionel Laurent, Annie Pruvost, Olivier |
author_sort |
Péréfarres, Frédéric |
title |
Real-time PCR for detection and quantification of Xanthomonas axonopodis pv. dieffenbachiae on Anthurium |
title_short |
Real-time PCR for detection and quantification of Xanthomonas axonopodis pv. dieffenbachiae on Anthurium |
title_full |
Real-time PCR for detection and quantification of Xanthomonas axonopodis pv. dieffenbachiae on Anthurium |
title_fullStr |
Real-time PCR for detection and quantification of Xanthomonas axonopodis pv. dieffenbachiae on Anthurium |
title_full_unstemmed |
Real-time PCR for detection and quantification of Xanthomonas axonopodis pv. dieffenbachiae on Anthurium |
title_sort |
real-time pcr for detection and quantification of xanthomonas axonopodis pv. dieffenbachiae on anthurium |
url |
http://agritrop.cirad.fr/547204/ |
work_keys_str_mv |
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