Diagnosis of Xanthomonas axonopodis pv. citri, causal agent of citrus canker, in commercial fruits by isolation and PCR-based methods
Aims: To show the results of the detection of an EU quarantine organism, Xanthomonas axonopodis pv. citri (Xac), in citrus fruits imported from countries where this bacterium is present, using an integrated approach that includes isolation, pathogenicity assays and molecular techniques. Methods and...
| Autores principales: | , , , , , |
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| Formato: | article |
| Lenguaje: | Inglés |
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Society for Applied Microbiology
2022
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| Materias: | |
| Acceso en línea: | http://hdl.handle.net/20.500.11939/8060 https://sfamjournals.onlinelibrary.wiley.com/doi/epdf/10.1111/j.1365-2672.2007.03484.x |
| _version_ | 1855032722028756992 |
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| author | Golmohammadi, Morteza Cubero, Jaime Penyalver, Jaime Quesada, Jose M. López, María M. Llop, Pablo |
| author_browse | Cubero, Jaime Golmohammadi, Morteza Llop, Pablo López, María M. Penyalver, Jaime Quesada, Jose M. |
| author_facet | Golmohammadi, Morteza Cubero, Jaime Penyalver, Jaime Quesada, Jose M. López, María M. Llop, Pablo |
| author_sort | Golmohammadi, Morteza |
| collection | ReDivia |
| description | Aims: To show the results of the detection of an EU quarantine organism, Xanthomonas axonopodis pv. citri (Xac), in citrus fruits imported from countries where this bacterium is present, using an integrated approach that includes isolation, pathogenicity assays and molecular techniques. Methods and Results: Citrus fruits with canker-like symptoms, exported to Spain from South American countries were analysed by several methods. Bacterial isolation, three conventional polymerase chain reaction (PCR) protocols, and real-time PCR with SYBR Green or a TaqMan probe, were compared. Canker-like lesions were disrupted in PBS buffer, and the extract used for bacterial isolation and DNA extraction followed by PCR amplification. Canker lesions, identified by PCR, showed viable bacteria in eleven of fifteen fruit samples. In 16 out of 130 lesions analysed from these samples, Xac was isolated, and pathogenicity on grapefruit leaves confirmed. By real-time PCR, using SYBR green or a Taqman probe, Xac was detected in 58 and 80 lesions respectively. By conventional PCR the bacterium was detected in 39–52 lesions depending on the protocol employed. Conclusions: An integrated approach for reliable detection of Xac in lesions of fruit samples, employing several techniques and with real-time PCR using a TaqMan probe as the fastest and most sensitive screening method, has been established and validated and is proposed as a useful tool for the analysis of Xac on fresh fruits. Significance and Impact of the Study: This work faces up to the real threat of the importation of citrus fruits that can harbour quarantine bacteria and will be useful in diagnostic laboratories for the analysis of commercial fresh fruits from countries where citrus canker is present. |
| format | article |
| id | ReDivia8060 |
| institution | Instituto Valenciano de Investigaciones Agrarias (IVIA) |
| language | Inglés |
| publishDate | 2022 |
| publishDateRange | 2022 |
| publishDateSort | 2022 |
| publisher | Society for Applied Microbiology |
| publisherStr | Society for Applied Microbiology |
| record_format | dspace |
| spelling | ReDivia80602025-04-25T14:48:40Z Diagnosis of Xanthomonas axonopodis pv. citri, causal agent of citrus canker, in commercial fruits by isolation and PCR-based methods Golmohammadi, Morteza Cubero, Jaime Penyalver, Jaime Quesada, Jose M. López, María M. Llop, Pablo Validations Real time-PCR Sensitivity Xanthomonas axonopodis H20 Plant diseases J10 Handling, transport, storage and protection of agricultural products E71 International trade protocols sensitivity Citrus fruits Diagnosis export Aims: To show the results of the detection of an EU quarantine organism, Xanthomonas axonopodis pv. citri (Xac), in citrus fruits imported from countries where this bacterium is present, using an integrated approach that includes isolation, pathogenicity assays and molecular techniques. Methods and Results: Citrus fruits with canker-like symptoms, exported to Spain from South American countries were analysed by several methods. Bacterial isolation, three conventional polymerase chain reaction (PCR) protocols, and real-time PCR with SYBR Green or a TaqMan probe, were compared. Canker-like lesions were disrupted in PBS buffer, and the extract used for bacterial isolation and DNA extraction followed by PCR amplification. Canker lesions, identified by PCR, showed viable bacteria in eleven of fifteen fruit samples. In 16 out of 130 lesions analysed from these samples, Xac was isolated, and pathogenicity on grapefruit leaves confirmed. By real-time PCR, using SYBR green or a Taqman probe, Xac was detected in 58 and 80 lesions respectively. By conventional PCR the bacterium was detected in 39–52 lesions depending on the protocol employed. Conclusions: An integrated approach for reliable detection of Xac in lesions of fruit samples, employing several techniques and with real-time PCR using a TaqMan probe as the fastest and most sensitive screening method, has been established and validated and is proposed as a useful tool for the analysis of Xac on fresh fruits. Significance and Impact of the Study: This work faces up to the real threat of the importation of citrus fruits that can harbour quarantine bacteria and will be useful in diagnostic laboratories for the analysis of commercial fresh fruits from countries where citrus canker is present. 2022-04-29T16:20:44Z 2022-04-29T16:20:44Z 2007 article publishedVersion Golmohammadi, M., Cubero, J., Peñalver, J., Quesada, J. M., López, M. M. & Llop, P. (2007). Diagnosis of Xanthomonas axonopodis pv. citri, causal agent of citrus canker, in commercial fruits by isolation and PCR‐based methods. Journal of Applied Microbiology, 103(6), 2309-2315. 1364-5072 http://hdl.handle.net/20.500.11939/8060 10.1111/j.1365-2672.2007.03484.x https://sfamjournals.onlinelibrary.wiley.com/doi/epdf/10.1111/j.1365-2672.2007.03484.x en info:eu-repo/grantAgreement/MEC/Programa Nacional de Recursos y Tecnologías Agroalimentarias/RTA2006-00149-00-00//Desarrollo de métodos de detección y estudios de supervivencia de Xanthomonas causantes de la cancrosis de los cítricos This research was supported by the agreement between the Ministerio de Agricultura of Spain, Subdirección General de Sanidad Vegetal and IVIA, through the project INIA RTA2006-00149-00-00, and by Plan de Vigilancia Fitosanitaria of the Generalitat Valenciana. Atribución-NoComercial-SinDerivadas 3.0 España http://creativecommons.org/licenses/by-nc-nd/3.0/es/ openAccess Society for Applied Microbiology electronico |
| spellingShingle | Validations Real time-PCR Sensitivity Xanthomonas axonopodis H20 Plant diseases J10 Handling, transport, storage and protection of agricultural products E71 International trade protocols sensitivity Citrus fruits Diagnosis export Golmohammadi, Morteza Cubero, Jaime Penyalver, Jaime Quesada, Jose M. López, María M. Llop, Pablo Diagnosis of Xanthomonas axonopodis pv. citri, causal agent of citrus canker, in commercial fruits by isolation and PCR-based methods |
| title | Diagnosis of Xanthomonas axonopodis pv. citri, causal agent of citrus canker, in commercial fruits by isolation and PCR-based methods |
| title_full | Diagnosis of Xanthomonas axonopodis pv. citri, causal agent of citrus canker, in commercial fruits by isolation and PCR-based methods |
| title_fullStr | Diagnosis of Xanthomonas axonopodis pv. citri, causal agent of citrus canker, in commercial fruits by isolation and PCR-based methods |
| title_full_unstemmed | Diagnosis of Xanthomonas axonopodis pv. citri, causal agent of citrus canker, in commercial fruits by isolation and PCR-based methods |
| title_short | Diagnosis of Xanthomonas axonopodis pv. citri, causal agent of citrus canker, in commercial fruits by isolation and PCR-based methods |
| title_sort | diagnosis of xanthomonas axonopodis pv citri causal agent of citrus canker in commercial fruits by isolation and pcr based methods |
| topic | Validations Real time-PCR Sensitivity Xanthomonas axonopodis H20 Plant diseases J10 Handling, transport, storage and protection of agricultural products E71 International trade protocols sensitivity Citrus fruits Diagnosis export |
| url | http://hdl.handle.net/20.500.11939/8060 https://sfamjournals.onlinelibrary.wiley.com/doi/epdf/10.1111/j.1365-2672.2007.03484.x |
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