Detection of DNA specific sequences of Spongospora subterranea in soil and potato tubers
A test has been developed for early identification of the casual agent for potato powdery scab (Spongospora subterranea fs subterranea). Identification was carried out in seeds and soil where this tuber is grown. A polymerase chain reaction (PCR) was set up for detecting 372, 390 and 391 bp ribosomal DNA internal transcribed spacer sequences (ITS) in the S. subterraneagenome. A method for extracting and purifying DNA from infected plant material (potato root nodes and pustules on the potato) was standardised. Plant tissue was obtained by potato tuber propagation using an inoculum from the pathogen in greenhouse conditions. After the PCR had been optimised and its sensitivity determined, a molecular methodology was validated by examining plant material infected with S. subterranea and soil samples infested with the pathogen obtained from the departments of Cundinamarca and Nariño. The PCR detected S. subterranea DNA from infected material and soil samples (all thirty samples from the experimental area analysed proved PCR positive). These results show that this molecular method was not just useful for the early detection of the pathogen in soil samples but as a tool for detecting or determining the possible presence of this micro-organism in places that have been declared f ree of S. subterranea and an effective form of quality control in producing the certif ied potato seed. Key words: Powdery scab, cystosori, internal transcribed spacer, PCR, plasmodiophorid.
Saved in:
| Main Authors: | , , |
|---|---|
| Format: | Digital revista |
| Language: | spa |
| Published: |
Universidad Nacional de Colombia - Sede Bogotá - Instituto de Biotecnología
2004
|
| Online Access: | https://revistas.unal.edu.co/index.php/biotecnologia/article/view/537 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| id |
oai:www.revistas.unal.edu.co:article-537 |
|---|---|
| record_format |
ojs |
| institution |
UNAL |
| collection |
OJS |
| country |
Colombia |
| countrycode |
CO |
| component |
Revista |
| access |
En linea |
| databasecode |
rev-biotecnologia-co |
| tag |
revista |
| region |
America del Sur |
| libraryname |
Sistema Nacional de Bibliotecas de la UNAL |
| language |
spa |
| format |
Digital |
| author |
Saavedra Rodríguez, Cristian Oswaldo Gómez González, Sandra Janeth Ángel Díaz, Jorge Evelio |
| spellingShingle |
Saavedra Rodríguez, Cristian Oswaldo Gómez González, Sandra Janeth Ángel Díaz, Jorge Evelio Detection of DNA specific sequences of Spongospora subterranea in soil and potato tubers |
| author_facet |
Saavedra Rodríguez, Cristian Oswaldo Gómez González, Sandra Janeth Ángel Díaz, Jorge Evelio |
| author_sort |
Saavedra Rodríguez, Cristian Oswaldo |
| title |
Detection of DNA specific sequences of Spongospora subterranea in soil and potato tubers |
| title_short |
Detection of DNA specific sequences of Spongospora subterranea in soil and potato tubers |
| title_full |
Detection of DNA specific sequences of Spongospora subterranea in soil and potato tubers |
| title_fullStr |
Detection of DNA specific sequences of Spongospora subterranea in soil and potato tubers |
| title_full_unstemmed |
Detection of DNA specific sequences of Spongospora subterranea in soil and potato tubers |
| title_sort |
detection of dna specific sequences of spongospora subterranea in soil and potato tubers |
| description |
A test has been developed for early identification of the casual agent for potato powdery scab (Spongospora subterranea fs subterranea). Identification was carried out in seeds and soil where this tuber is grown. A polymerase chain reaction (PCR) was set up for detecting 372, 390 and 391 bp ribosomal DNA internal transcribed spacer sequences (ITS) in the S. subterraneagenome. A method for extracting and purifying DNA from infected plant material (potato root nodes and pustules on the potato) was standardised. Plant tissue was obtained by potato tuber propagation using an inoculum from the pathogen in greenhouse conditions. After the PCR had been optimised and its sensitivity determined, a molecular methodology was validated by examining plant material infected with S. subterranea and soil samples infested with the pathogen obtained from the departments of Cundinamarca and Nariño. The PCR detected S. subterranea DNA from infected material and soil samples (all thirty samples from the experimental area analysed proved PCR positive). These results show that this molecular method was not just useful for the early detection of the pathogen in soil samples but as a tool for detecting or determining the possible presence of this micro-organism in places that have been declared f ree of S. subterranea and an effective form of quality control in producing the certif ied potato seed.
Key words: Powdery scab, cystosori, internal transcribed spacer, PCR, plasmodiophorid. |
| publisher |
Universidad Nacional de Colombia - Sede Bogotá - Instituto de Biotecnología |
| publishDate |
2004 |
| url |
https://revistas.unal.edu.co/index.php/biotecnologia/article/view/537 |
| work_keys_str_mv |
AT saavedrarodriguezcristianoswaldo detectionofdnaspecificsequencesofspongosporasubterraneainsoilandpotatotubers AT gomezgonzalezsandrajaneth detectionofdnaspecificsequencesofspongosporasubterraneainsoilandpotatotubers AT angeldiazjorgeevelio detectionofdnaspecificsequencesofspongosporasubterraneainsoilandpotatotubers AT saavedrarodriguezcristianoswaldo detecciondesecuenciasespecificasdeadndespongosporasubterraneaensueloytuberculosdepapa AT gomezgonzalezsandrajaneth detecciondesecuenciasespecificasdeadndespongosporasubterraneaensueloytuberculosdepapa AT angeldiazjorgeevelio detecciondesecuenciasespecificasdeadndespongosporasubterraneaensueloytuberculosdepapa |
| _version_ |
1794799475561070592 |
| spelling |
oai:www.revistas.unal.edu.co:article-5372007-05-03T17:23:13Z Detection of DNA specific sequences of Spongospora subterranea in soil and potato tubers Detección de secuencias específicas de ADN de Spongospora subterranea en suelo y tubérculos de papa Saavedra Rodríguez, Cristian Oswaldo Gómez González, Sandra Janeth Ángel Díaz, Jorge Evelio sarna polvosa quistosoros espaciador de transcripción interna PCR plasmodiofóridos V A test has been developed for early identification of the casual agent for potato powdery scab (Spongospora subterranea fs subterranea). Identification was carried out in seeds and soil where this tuber is grown. A polymerase chain reaction (PCR) was set up for detecting 372, 390 and 391 bp ribosomal DNA internal transcribed spacer sequences (ITS) in the S. subterraneagenome. A method for extracting and purifying DNA from infected plant material (potato root nodes and pustules on the potato) was standardised. Plant tissue was obtained by potato tuber propagation using an inoculum from the pathogen in greenhouse conditions. After the PCR had been optimised and its sensitivity determined, a molecular methodology was validated by examining plant material infected with S. subterranea and soil samples infested with the pathogen obtained from the departments of Cundinamarca and Nariño. The PCR detected S. subterranea DNA from infected material and soil samples (all thirty samples from the experimental area analysed proved PCR positive). These results show that this molecular method was not just useful for the early detection of the pathogen in soil samples but as a tool for detecting or determining the possible presence of this micro-organism in places that have been declared f ree of S. subterranea and an effective form of quality control in producing the certif ied potato seed. Key words: Powdery scab, cystosori, internal transcribed spacer, PCR, plasmodiophorid. Con el fin de identificar de manera precoz el agente causal de la sarna polvosa de la papa (Spongospora subterranea fs subterranea) tanto en semillas como en suelos aptos para el cultivo de este tubérculo, se ha desarrollado una prueba basada en la reacción en cadena de la polimerasa (PCR) para la detección de tres secuencias ITS (espaciadores de transcripción interna específicos del ADNr) de 372,390 y 391 pb presentes en el genoma de S. subterranea. Para ello se estandarizó una metodología de extracción y purificación de ADN del microorganismo a partir de tejido vegetal infectado (agallas de raíz de papa y pústulas en papa) obtenido en pruebas de propagación del patógeno en invernadero. Una vez optimizada la PCR y determinada su sensibilidad, se validó la metodología molecular examinando muestras de tejido vegetal infectado y suelo infestado por el patógeno, provenientes de los departamentos de Cundinamarca y Nariño. La PCR detectó ADN del microorganismo tanto en el material vegetal infectado, como en las muestras de suelo analizadas (las 30 submuestras del área experimental fueron PCR positivas). Estos resultados muestran que dicha metodología se presenta no sólo como una técnica útil para la detección rápida del patógeno en suelo, sino también como una herramienta para determinar la presencia del microorganismo en suelos declarados libres de éste y como una alternativa eficaz para el control de calidad en la producción de semilla certificada de papa libre del patógeno. Palabras clave: sarna polvosa, quistosoros, espaciador de transcripción interna, PCR, plasmodiofóridos. Universidad Nacional de Colombia - Sede Bogotá - Instituto de Biotecnología 2004-01-01 info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion Artículo revisado por pares application/pdf application/msword text/html https://revistas.unal.edu.co/index.php/biotecnologia/article/view/537 Revista Colombiana de Biotecnología; Vol. 6 No. 1 (2004); 14-23 Revista Colombiana de Biotecnología; Vol. 6 Núm. 1 (2004); 14-23 1909-8758 0123-3475 spa https://revistas.unal.edu.co/index.php/biotecnologia/article/view/537/1028 https://revistas.unal.edu.co/index.php/biotecnologia/article/view/537/1029 https://revistas.unal.edu.co/index.php/biotecnologia/article/view/537/1030 Derechos de autor 2004 Revista Colombiana de Biotecnología https://creativecommons.org/licenses/by/4.0 |