A real-time PCR quantitative detection assay for Pseudomonas savastanoi pv. nerii in Nerium oleander
Autor: | P. Bella, G. Licciardello, M. Tessitori, V. Catara |
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Jazyk: | angličtina |
Rok vydání: | 2009 |
Předmět: | |
Zdroj: | Phytopathologia Mediterranea, Vol 47, Iss 3 (2009) |
Druh dokumentu: | article |
ISSN: | 0031-9465 1593-2095 |
DOI: | 10.14601/Phytopathol_Mediterr-2724 |
Popis: | A real-time PCR assay based on TaqMan chemistry was developed for the detection of Pseudomonas savastanoi pathovars that cause bacterial knot disease on different plant species. Primers and probe sequences were based on the iaaL gene coding for (indole-3-acetyl)-L-lysine synthetase and previously used in conventional PCR tests. Assay specificity was tested with an extended range of strains of P. savastanoi from eight hosts, with 13 other Pseudomonas spp., and with other microorganisms naturally occurring on or in oleander plants. A pure culture cell suspension was quantifi ed over a seven log concentration range (108 to 102 cfu ml-1 ). Different protocols were developed for the detection and quantifi cation of P. savastanoi pv. nerii from symptomatic and asymptomatic oleander plants. A 24-h bacterial enrichment step either on PVF-1 or OKA-M broth improved the sensitivity of the assay, making it suitable to screen planting material for latent infections. |
Databáze: | Directory of Open Access Journals |
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