Development of A Nested-MultiLocus Sequence Typing Approach for A Highly Sensitive and Specific Identification of Xylella fastidiosa Subspecies Directly from Plant Samples
© 2020 by the authors.
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| Format: | artículo biblioteca |
| Language: | English |
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Multidisciplinary Digital Publishing Institute
2020
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| Subjects: | Xylella fastidiosa, Direct typing technique, MLST, Nested PCR, |
| Online Access: | http://dx.doi.org/10.13039/501100009599 http://dx.doi.org/10.13039/501100003359 http://dx.doi.org/10.13039/501100000780 http://dx.doi.org/10.13039/501100003339 |
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dig-ias-es-10261-2184022020-10-08T01:14:16Z Development of A Nested-MultiLocus Sequence Typing Approach for A Highly Sensitive and Specific Identification of Xylella fastidiosa Subspecies Directly from Plant Samples Cesbron, Sophie Dupas, Enora Beaurepère, Quentin Briand, Martial Montes Borrego, Miguel Velasco-Amo, María Pilar Landa, Blanca B. Jacques, M. A. European Commission Interprofesional del Aceite de Oliva Español Consejo Superior de Investigaciones Científicas (España) Generalitat Valenciana Ministerio de Agricultura, Pesca y Alimentación y Medio Ambiente (España) European Commission Xylella fastidiosa Direct typing technique MLST Nested PCR © 2020 by the authors. Identification of sequence types (ST) of Xylella fastidiosa based on direct MultiLocus Sequence Typing (MLST) of plant DNA samples is partly efficient. In order to improve the sensitivity of X. fastidiosa identification, we developed a direct nested-MLST assay on plant extracted DNA. This method was performed based on a largely used scheme targeting seven housekeeping gene (HKG) loci (cysG, gltT, holC, leuA, malF, nuoL, petC). Samples analyzed included 49 plant species and two insect species (Philaenus spumarius, Neophilaenus campestris) that were collected in 2017 (106 plant samples in France), in 2018 (162 plant samples in France, 40 plant samples and 26 insect samples in Spain), and in 2019 (30 plant samples in Spain). With the nested approach, a significant higher number of samples were amplified. The threshold was improved by 100 to 1000 times compared to conventional PCR. Using nested-MLST assay, plants that were not yet considered hosts tested positive and revealed novel alleles in France, whereas for Spanish samples it was possible to assign the subspecies or ST to samples considered as new hosts in Europe. Direct typing by nested-MLST from plant material has an increased sensitivity and may be useful for epidemiological purposes. ED salary was funded by INRA SPE division and Anses. This work received support from the European Union’s Horizon 2020 research and innovation program under grant agreement 727987 XF-ACTORS (Xylella fastidiosa Active Containment Through a multidisciplinary-Oriented Research Strategy), and from Projects E-RTA2017-00004-C06-02 from ‘Programa Estatal de I+D Orientada a los Retos de la Sociedad’ from Spanish State Research Agency and from the Organización Interprofesional del Aceite de Oliva Español, CSIC Intramural Project 2018 40E111, and from “Conselleria de Agricultura, Desarrollo Rural, Emergencia Climática y Transición Ecológica” from Valencia region, and the Ministry of Agriculture, Fisheries and Food of Spain. The present work reflects only the authors’view and the EU funding agency is not responsible for any use that may be made of the information it contains. Peer reviewed 2020-08-21T13:49:34Z 2020-08-21T13:49:34Z 2020 2020-08-21T13:49:35Z artículo http://purl.org/coar/resource_type/c_6501 Agronomy 10(8): 1099 (2020) hdl.handle.net/10261/218402 10.3390/agronomy10081099 2073-4395 http://dx.doi.org/10.13039/501100009599 http://dx.doi.org/10.13039/501100003359 http://dx.doi.org/10.13039/501100000780 http://dx.doi.org/10.13039/501100003339 en #PLACEHOLDER_PARENT_METADATA_VALUE# info:eu-repo/grantAgreement/EC/H2020/727987 Publisher’s version https://doi.org/10.3390/agronomy10081099 Sí open Multidisciplinary Digital Publishing Institute |
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Xylella fastidiosa Direct typing technique MLST Nested PCR Xylella fastidiosa Direct typing technique MLST Nested PCR |
| spellingShingle |
Xylella fastidiosa Direct typing technique MLST Nested PCR Xylella fastidiosa Direct typing technique MLST Nested PCR Cesbron, Sophie Dupas, Enora Beaurepère, Quentin Briand, Martial Montes Borrego, Miguel Velasco-Amo, María Pilar Landa, Blanca B. Jacques, M. A. Development of A Nested-MultiLocus Sequence Typing Approach for A Highly Sensitive and Specific Identification of Xylella fastidiosa Subspecies Directly from Plant Samples |
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© 2020 by the authors. |
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European Commission |
| author_facet |
European Commission Cesbron, Sophie Dupas, Enora Beaurepère, Quentin Briand, Martial Montes Borrego, Miguel Velasco-Amo, María Pilar Landa, Blanca B. Jacques, M. A. |
| format |
artículo |
| topic_facet |
Xylella fastidiosa Direct typing technique MLST Nested PCR |
| author |
Cesbron, Sophie Dupas, Enora Beaurepère, Quentin Briand, Martial Montes Borrego, Miguel Velasco-Amo, María Pilar Landa, Blanca B. Jacques, M. A. |
| author_sort |
Cesbron, Sophie |
| title |
Development of A Nested-MultiLocus Sequence Typing Approach for A Highly Sensitive and Specific Identification of Xylella fastidiosa Subspecies Directly from Plant Samples |
| title_short |
Development of A Nested-MultiLocus Sequence Typing Approach for A Highly Sensitive and Specific Identification of Xylella fastidiosa Subspecies Directly from Plant Samples |
| title_full |
Development of A Nested-MultiLocus Sequence Typing Approach for A Highly Sensitive and Specific Identification of Xylella fastidiosa Subspecies Directly from Plant Samples |
| title_fullStr |
Development of A Nested-MultiLocus Sequence Typing Approach for A Highly Sensitive and Specific Identification of Xylella fastidiosa Subspecies Directly from Plant Samples |
| title_full_unstemmed |
Development of A Nested-MultiLocus Sequence Typing Approach for A Highly Sensitive and Specific Identification of Xylella fastidiosa Subspecies Directly from Plant Samples |
| title_sort |
development of a nested-multilocus sequence typing approach for a highly sensitive and specific identification of xylella fastidiosa subspecies directly from plant samples |
| publisher |
Multidisciplinary Digital Publishing Institute |
| publishDate |
2020 |
| url |
http://dx.doi.org/10.13039/501100009599 http://dx.doi.org/10.13039/501100003359 http://dx.doi.org/10.13039/501100000780 http://dx.doi.org/10.13039/501100003339 |
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