A simple extraction procedure for efficient routine detection of pathogenic bacteria in plant material by polymerase chain reaction
A simple and rapid method for extracting DNA from plants based on the use of an extraction buffer and precipitation with isopropanol was assayed to see its usefulness in detecting pathogenic bacteria in plant material. The method was compared with a phenol-chloroform standard procedure obtaining hig...
| Autores principales: | , , , , |
|---|---|
| Formato: | Artículo |
| Lenguaje: | Inglés |
| Publicado: |
2017
|
| Acceso en línea: | http://hdl.handle.net/20.500.11939/5497 https://www.sciencedirect.com/science/article/abs/pii/S0167701299000330?via%3Dihub |
| _version_ | 1855491935069798400 |
|---|---|
| author | Llop, Pablo Caruso, Paola Cubero, Jaime Morente, Clara López, María M. |
| author_browse | Caruso, Paola Cubero, Jaime Llop, Pablo López, María M. Morente, Clara |
| author_facet | Llop, Pablo Caruso, Paola Cubero, Jaime Morente, Clara López, María M. |
| author_sort | Llop, Pablo |
| collection | ReDivia |
| description | A simple and rapid method for extracting DNA from plants based on the use of an extraction buffer and precipitation with isopropanol was assayed to see its usefulness in detecting pathogenic bacteria in plant material. The method was compared with a phenol-chloroform standard procedure obtaining higher sensitivity levels of detection. The protocol developed was efficient for detecting a Gram-positive bacterium, Clavibacter michiganensis subsp. sepedonicus and several Gram-negative pathogenic bacteria (Ralstonia solanacearum, Erwinia amylovora, Xanthomonas axonopodis pv. citri) with a sensitivity of 10(2)-10(3) cfu/ml in spiked samples. It was also efficient to specifically identify such bacteria in naturally infected plant material. This procedure is proposed as a routine tool for detection of plant pathogenic bacteria, as well as in environmental microbiology and biotechnology studies. |
| format | Artículo |
| id | ReDivia5497 |
| institution | Instituto Valenciano de Investigaciones Agrarias (IVIA) |
| language | Inglés |
| publishDate | 2017 |
| publishDateRange | 2017 |
| publishDateSort | 2017 |
| record_format | dspace |
| spelling | ReDivia54972025-04-25T14:43:07Z A simple extraction procedure for efficient routine detection of pathogenic bacteria in plant material by polymerase chain reaction Llop, Pablo Caruso, Paola Cubero, Jaime Morente, Clara López, María M. A simple and rapid method for extracting DNA from plants based on the use of an extraction buffer and precipitation with isopropanol was assayed to see its usefulness in detecting pathogenic bacteria in plant material. The method was compared with a phenol-chloroform standard procedure obtaining higher sensitivity levels of detection. The protocol developed was efficient for detecting a Gram-positive bacterium, Clavibacter michiganensis subsp. sepedonicus and several Gram-negative pathogenic bacteria (Ralstonia solanacearum, Erwinia amylovora, Xanthomonas axonopodis pv. citri) with a sensitivity of 10(2)-10(3) cfu/ml in spiked samples. It was also efficient to specifically identify such bacteria in naturally infected plant material. This procedure is proposed as a routine tool for detection of plant pathogenic bacteria, as well as in environmental microbiology and biotechnology studies. 2017-06-01T10:12:27Z 2017-06-01T10:12:27Z 1999 JUL 1999 article Llop, P., Caruso, P., Cubero, J., Morente, C. & Lopez, M. M. (1999). A simple extraction procedure for efficient routine detection of pathogenic bacteria in plant material by polymerase chain reaction. Journal of microbiological methods, 37(1), 23-31. 0167-7012 http://hdl.handle.net/20.500.11939/5497 10.1016/S0167-7012(99)00033-0 https://www.sciencedirect.com/science/article/abs/pii/S0167701299000330?via%3Dihub en openAccess Impreso |
| spellingShingle | Llop, Pablo Caruso, Paola Cubero, Jaime Morente, Clara López, María M. A simple extraction procedure for efficient routine detection of pathogenic bacteria in plant material by polymerase chain reaction |
| title | A simple extraction procedure for efficient routine detection of pathogenic bacteria in plant material by polymerase chain reaction |
| title_full | A simple extraction procedure for efficient routine detection of pathogenic bacteria in plant material by polymerase chain reaction |
| title_fullStr | A simple extraction procedure for efficient routine detection of pathogenic bacteria in plant material by polymerase chain reaction |
| title_full_unstemmed | A simple extraction procedure for efficient routine detection of pathogenic bacteria in plant material by polymerase chain reaction |
| title_short | A simple extraction procedure for efficient routine detection of pathogenic bacteria in plant material by polymerase chain reaction |
| title_sort | simple extraction procedure for efficient routine detection of pathogenic bacteria in plant material by polymerase chain reaction |
| url | http://hdl.handle.net/20.500.11939/5497 https://www.sciencedirect.com/science/article/abs/pii/S0167701299000330?via%3Dihub |
| work_keys_str_mv | AT lloppablo asimpleextractionprocedureforefficientroutinedetectionofpathogenicbacteriainplantmaterialbypolymerasechainreaction AT carusopaola asimpleextractionprocedureforefficientroutinedetectionofpathogenicbacteriainplantmaterialbypolymerasechainreaction AT cuberojaime asimpleextractionprocedureforefficientroutinedetectionofpathogenicbacteriainplantmaterialbypolymerasechainreaction AT morenteclara asimpleextractionprocedureforefficientroutinedetectionofpathogenicbacteriainplantmaterialbypolymerasechainreaction AT lopezmariam asimpleextractionprocedureforefficientroutinedetectionofpathogenicbacteriainplantmaterialbypolymerasechainreaction AT lloppablo simpleextractionprocedureforefficientroutinedetectionofpathogenicbacteriainplantmaterialbypolymerasechainreaction AT carusopaola simpleextractionprocedureforefficientroutinedetectionofpathogenicbacteriainplantmaterialbypolymerasechainreaction AT cuberojaime simpleextractionprocedureforefficientroutinedetectionofpathogenicbacteriainplantmaterialbypolymerasechainreaction AT morenteclara simpleextractionprocedureforefficientroutinedetectionofpathogenicbacteriainplantmaterialbypolymerasechainreaction AT lopezmariam simpleextractionprocedureforefficientroutinedetectionofpathogenicbacteriainplantmaterialbypolymerasechainreaction |