Detection of Ralstonia solanacearum (Rs) in latently infected potato stem extracts by post-enrichment qPCR.
A sensitive method for detection of Ralstonia solanacearum in latently infected potato stem extracts has been achieved by previous enrichment procedure followed by Real time PCR (qPCR). Sensitivity of qPCR before and after enrichment (48h of incubation of the stem extract with modified SMSA broth at...
| Autores principales: | , , |
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| Formato: | Conference Paper |
| Lenguaje: | Inglés |
| Publicado: |
2012
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| Materias: | |
| Acceso en línea: | https://hdl.handle.net/10568/66293 |
| _version_ | 1855541670768017408 |
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| author | Gutarra, L. Fuentes, S. Kreuze, Jan F. |
| author_browse | Fuentes, S. Gutarra, L. Kreuze, Jan F. |
| author_facet | Gutarra, L. Fuentes, S. Kreuze, Jan F. |
| author_sort | Gutarra, L. |
| collection | Repository of Agricultural Research Outputs (CGSpace) |
| description | A sensitive method for detection of Ralstonia solanacearum in latently infected potato stem extracts has been achieved by previous enrichment procedure followed by Real time PCR (qPCR). Sensitivity of qPCR before and after enrichment (48h of incubation of the stem extract with modified SMSA broth at 30oC) was compared with other techniques such as NCM-ELISA and R.solanacearum isolation in Kelman’s medium. Before enrichment procedure, 174.6 cells/ml were detected in Kelman’s medium, 1.71 x 106 cells/ml by NCM-ELISA and 1.29x105 cells/ by qPCR. After enrichment, sensitivities of post enrichment qPCR, NCM- ELISA and isolation on modified Kelman’s medium were similar. As few as 6.56 cells/ml were detected in latently infected potato stem extracts. Serial dilutions of naturally-infected stem extract before enrichment allowed the quantification of populations of R. solanacearum in each stem extract. Post –enrichment qPCR combines the advantages of high sensitivity, ease and speed, but it requires expensive laboratory equipment. Thus it can be used by seed and breeding programmes in developing countries only to confirm positive results obtained by serological tests used for seed quality control and for assessing susceptibility of breeding lines to bacterial wilt. |
| format | Conference Paper |
| id | CGSpace66293 |
| institution | CGIAR Consortium |
| language | Inglés |
| publishDate | 2012 |
| publishDateRange | 2012 |
| publishDateSort | 2012 |
| record_format | dspace |
| spelling | CGSpace662932025-11-06T14:04:12Z Detection of Ralstonia solanacearum (Rs) in latently infected potato stem extracts by post-enrichment qPCR. Gutarra, L. Fuentes, S. Kreuze, Jan F. potatoes ralstonia solanacearum bacterial diseases plant extracts plant diseases latent infections pcr methods A sensitive method for detection of Ralstonia solanacearum in latently infected potato stem extracts has been achieved by previous enrichment procedure followed by Real time PCR (qPCR). Sensitivity of qPCR before and after enrichment (48h of incubation of the stem extract with modified SMSA broth at 30oC) was compared with other techniques such as NCM-ELISA and R.solanacearum isolation in Kelman’s medium. Before enrichment procedure, 174.6 cells/ml were detected in Kelman’s medium, 1.71 x 106 cells/ml by NCM-ELISA and 1.29x105 cells/ by qPCR. After enrichment, sensitivities of post enrichment qPCR, NCM- ELISA and isolation on modified Kelman’s medium were similar. As few as 6.56 cells/ml were detected in latently infected potato stem extracts. Serial dilutions of naturally-infected stem extract before enrichment allowed the quantification of populations of R. solanacearum in each stem extract. Post –enrichment qPCR combines the advantages of high sensitivity, ease and speed, but it requires expensive laboratory equipment. Thus it can be used by seed and breeding programmes in developing countries only to confirm positive results obtained by serological tests used for seed quality control and for assessing susceptibility of breeding lines to bacterial wilt. 2012 2015-05-19T10:56:24Z 2015-05-19T10:56:24Z Conference Paper https://hdl.handle.net/10568/66293 en Open Access application/pdf Gutarra, L.; Segundo, F.; Kreuze, J. 2012. Detection of Ralstonia solanacearum (Rs) in latently infected potato stem extracts by post-enrichment qPCR. Memorias de la 58ava reunión anual: Programas y resumenes. 58. Annual Meeting of The Interamerican Society for Tropical Horticulture (ISTH). 16. Congreso de la Sociedad Peruana de Horticultura. Lima (Perú). 3 - 6 Sep 2012. Lima (Peru). ISTH Universidad Nacional Agraria La Molina (UNALM). p. 64. Abstract |
| spellingShingle | potatoes ralstonia solanacearum bacterial diseases plant extracts plant diseases latent infections pcr methods Gutarra, L. Fuentes, S. Kreuze, Jan F. Detection of Ralstonia solanacearum (Rs) in latently infected potato stem extracts by post-enrichment qPCR. |
| title | Detection of Ralstonia solanacearum (Rs) in latently infected potato stem extracts by post-enrichment qPCR. |
| title_full | Detection of Ralstonia solanacearum (Rs) in latently infected potato stem extracts by post-enrichment qPCR. |
| title_fullStr | Detection of Ralstonia solanacearum (Rs) in latently infected potato stem extracts by post-enrichment qPCR. |
| title_full_unstemmed | Detection of Ralstonia solanacearum (Rs) in latently infected potato stem extracts by post-enrichment qPCR. |
| title_short | Detection of Ralstonia solanacearum (Rs) in latently infected potato stem extracts by post-enrichment qPCR. |
| title_sort | detection of ralstonia solanacearum rs in latently infected potato stem extracts by post enrichment qpcr |
| topic | potatoes ralstonia solanacearum bacterial diseases plant extracts plant diseases latent infections pcr methods |
| url | https://hdl.handle.net/10568/66293 |
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