Improvement of three nucleic acid isolation protocols for an overall diagnosis of viruses on six vegetative propagated plants
Biological Resources Center (BRCs) must be able to guarantee the sanitary status of the resources they distribute, in order to prevent the spread or emergence of diseases. However, BRCs' vegetatively propagated crops do not benefit from the partial sanitation occurring through a seed cycle. This is particularly a problem for viral diseases, which have an overall high prevalence in vegetatively propagated crops. Variously effective sanitation methods exist for recovering virus-free plants but their successful implementation depends on on the availability of sensitive, polyvalent and reliable diagnosis tests for all relevant virus species. The main objective of the SafePGR project is to improve the knowledge of the diversity of viruses infecting the vegetatively propagated crops addressed by the partners' BRCs (Universidad do Açores, Universidad da Madeira, INRA-CIRAD Guadeloupe and CIRAD La Réunion). Among the various issues addressed in achieving the goals of the SafePGR project, we need to develop new tools for an overall diagnosis of viruses. Thus, recent metagenomics methods associated with high-throughput sequencing will be tested. For this purpose, we started to develop and adapt three different nucleic acids extractions on six plants species: banana, garlic, sugarcane, sweet potato, vanilla and yam. First, we succeeded to extract small RNAs using Trizol or phenol:chloroform methods on these six species. Then, we have developed a protocol to semi-purify viral particles. The third protocol consisted in an enrichment of double-stranded RNAs. The quality and quantity of extracted nucleic acid varied among plant species. Overall, the extracted RNAs from garlic, sugarcane, sweet potato and vanilla were fulfilling criteria of quality and quantity for being used for metagenomic approaches whereas the ones from banana and yam were not adequate. These preliminary results tend to indicate that it would be probably difficult to develop a universal nucleic acid isolation method that could be routinely used by our partners' BRCs. (Texte intégral)
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dig-cirad-fr-5680372022-05-24T16:03:07Z http://agritrop.cirad.fr/568037/ http://agritrop.cirad.fr/568037/ Improvement of three nucleic acid isolation protocols for an overall diagnosis of viruses on six vegetative propagated plants. Julian Charlotte, Bernardo Pauline, Fernandez Emmanuel, Galzi Serge, Grisoni Michel, Da Silva Mendoça D.M., Pavis Claudie, Roumagnac Philippe, Filloux Denis. 2013. In : 14èmes Rencontres de virologie végétale (RVV 2013) : Aussois, France, 13-17 janvier 2013. Marais Armelle (ed.), Revers Frédéric (ed.). SFP, INRA. Paris : SFP, Résumé, 75. Rencontres de Virologie Végétale. 14, Aussois, France, 13 Janvier 2013/17 Janvier 2013. Researchers Improvement of three nucleic acid isolation protocols for an overall diagnosis of viruses on six vegetative propagated plants Julian, Charlotte Bernardo, Pauline Fernandez, Emmanuel Galzi, Serge Grisoni, Michel Da Silva Mendoça, D.M. Pavis, Claudie Roumagnac, Philippe Filloux, Denis eng 2013 SFP 14èmes Rencontres de virologie végétale (RVV 2013) : Aussois, France, 13-17 janvier 2013 H20 - Maladies des plantes F02 - Multiplication végétative des plantes Biological Resources Center (BRCs) must be able to guarantee the sanitary status of the resources they distribute, in order to prevent the spread or emergence of diseases. However, BRCs' vegetatively propagated crops do not benefit from the partial sanitation occurring through a seed cycle. This is particularly a problem for viral diseases, which have an overall high prevalence in vegetatively propagated crops. Variously effective sanitation methods exist for recovering virus-free plants but their successful implementation depends on on the availability of sensitive, polyvalent and reliable diagnosis tests for all relevant virus species. The main objective of the SafePGR project is to improve the knowledge of the diversity of viruses infecting the vegetatively propagated crops addressed by the partners' BRCs (Universidad do Açores, Universidad da Madeira, INRA-CIRAD Guadeloupe and CIRAD La Réunion). Among the various issues addressed in achieving the goals of the SafePGR project, we need to develop new tools for an overall diagnosis of viruses. Thus, recent metagenomics methods associated with high-throughput sequencing will be tested. For this purpose, we started to develop and adapt three different nucleic acids extractions on six plants species: banana, garlic, sugarcane, sweet potato, vanilla and yam. First, we succeeded to extract small RNAs using Trizol or phenol:chloroform methods on these six species. Then, we have developed a protocol to semi-purify viral particles. The third protocol consisted in an enrichment of double-stranded RNAs. The quality and quantity of extracted nucleic acid varied among plant species. Overall, the extracted RNAs from garlic, sugarcane, sweet potato and vanilla were fulfilling criteria of quality and quantity for being used for metagenomic approaches whereas the ones from banana and yam were not adequate. These preliminary results tend to indicate that it would be probably difficult to develop a universal nucleic acid isolation method that could be routinely used by our partners' BRCs. (Texte intégral) conference_item info:eu-repo/semantics/conferenceObject Conference info:eu-repo/semantics/publishedVersion http://agritrop.cirad.fr/568037/1/document_568037.pdf application/pdf Cirad license info:eu-repo/semantics/openAccess https://agritrop.cirad.fr/mention_legale.html http://catalogue-bibliotheques.cirad.fr/cgi-bin/koha/opac-detail.pl?biblionumber=214172 |
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H20 - Maladies des plantes F02 - Multiplication végétative des plantes H20 - Maladies des plantes F02 - Multiplication végétative des plantes |
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H20 - Maladies des plantes F02 - Multiplication végétative des plantes H20 - Maladies des plantes F02 - Multiplication végétative des plantes Julian, Charlotte Bernardo, Pauline Fernandez, Emmanuel Galzi, Serge Grisoni, Michel Da Silva Mendoça, D.M. Pavis, Claudie Roumagnac, Philippe Filloux, Denis Improvement of three nucleic acid isolation protocols for an overall diagnosis of viruses on six vegetative propagated plants |
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Biological Resources Center (BRCs) must be able to guarantee the sanitary status of the resources they distribute, in order to prevent the spread or emergence of diseases. However, BRCs' vegetatively propagated crops do not benefit from the partial sanitation occurring through a seed cycle. This is particularly a problem for viral diseases, which have an overall high prevalence in vegetatively propagated crops. Variously effective sanitation methods exist for recovering virus-free plants but their successful implementation depends on on the availability of sensitive, polyvalent and reliable diagnosis tests for all relevant virus species. The main objective of the SafePGR project is to improve the knowledge of the diversity of viruses infecting the vegetatively propagated crops addressed by the partners' BRCs (Universidad do Açores, Universidad da Madeira, INRA-CIRAD Guadeloupe and CIRAD La Réunion). Among the various issues addressed in achieving the goals of the SafePGR project, we need to develop new tools for an overall diagnosis of viruses. Thus, recent metagenomics methods associated with high-throughput sequencing will be tested. For this purpose, we started to develop and adapt three different nucleic acids extractions on six plants species: banana, garlic, sugarcane, sweet potato, vanilla and yam. First, we succeeded to extract small RNAs using Trizol or phenol:chloroform methods on these six species. Then, we have developed a protocol to semi-purify viral particles. The third protocol consisted in an enrichment of double-stranded RNAs. The quality and quantity of extracted nucleic acid varied among plant species. Overall, the extracted RNAs from garlic, sugarcane, sweet potato and vanilla were fulfilling criteria of quality and quantity for being used for metagenomic approaches whereas the ones from banana and yam were not adequate. These preliminary results tend to indicate that it would be probably difficult to develop a universal nucleic acid isolation method that could be routinely used by our partners' BRCs. (Texte intégral) |
format |
conference_item |
topic_facet |
H20 - Maladies des plantes F02 - Multiplication végétative des plantes |
author |
Julian, Charlotte Bernardo, Pauline Fernandez, Emmanuel Galzi, Serge Grisoni, Michel Da Silva Mendoça, D.M. Pavis, Claudie Roumagnac, Philippe Filloux, Denis |
author_facet |
Julian, Charlotte Bernardo, Pauline Fernandez, Emmanuel Galzi, Serge Grisoni, Michel Da Silva Mendoça, D.M. Pavis, Claudie Roumagnac, Philippe Filloux, Denis |
author_sort |
Julian, Charlotte |
title |
Improvement of three nucleic acid isolation protocols for an overall diagnosis of viruses on six vegetative propagated plants |
title_short |
Improvement of three nucleic acid isolation protocols for an overall diagnosis of viruses on six vegetative propagated plants |
title_full |
Improvement of three nucleic acid isolation protocols for an overall diagnosis of viruses on six vegetative propagated plants |
title_fullStr |
Improvement of three nucleic acid isolation protocols for an overall diagnosis of viruses on six vegetative propagated plants |
title_full_unstemmed |
Improvement of three nucleic acid isolation protocols for an overall diagnosis of viruses on six vegetative propagated plants |
title_sort |
improvement of three nucleic acid isolation protocols for an overall diagnosis of viruses on six vegetative propagated plants |
publisher |
SFP |
url |
http://agritrop.cirad.fr/568037/ http://agritrop.cirad.fr/568037/1/document_568037.pdf |
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