Optimization of a real-time PCR assay for the detection of the quarantine pathogen Melampsora medusae f. sp. deltoidae
| Autor: | Richard C. Hamelin, Cécile Guinet, Anne-Laure Boutigny, Agathe Vialle, Axelle Andrieux, Pascal Frey, Renaud Ioos, Claude Husson |
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| Přispěvatelé: | Ecosystèmes Forestiers, Agroressources, Bioprocédés et Alimentation (EFABA), Université de Lorraine (UL), Canadian Forest Service - CFS (CANADA), Interactions Arbres-Microorganismes (IAM), Université de Lorraine (UL)-Institut National de la Recherche Agronomique (INRA), Institut National de la Recherche Agronomique (INRA)-Université de Lorraine (UL) |
| Rok vydání: | 2012 |
| Předmět: |
0106 biological sciences
[SDV]Life Sciences [q-bio] Melampsora Mycology Real-Time Polymerase Chain Reaction 01 natural sciences Sensitivity and Specificity Microbiology law.invention 03 medical and health sciences Quarantine pathogen law Botany RNA Ribosomal 28S Genetics Ecology Evolution Behavior and Systematics Polymerase chain reaction 030304 developmental biology Urediniospore Phytosanitary certification DNA Primers Plant Diseases 0303 health sciences biology Basidiomycota fungi Reproducibility of Results RNA Fungal Ribosomal RNA biology.organism_classification 3. Good health Europe Infectious Diseases Real-time polymerase chain reaction Populus Quarantine Melampsora medusae PEST analysis Poplar rust Oligonucleotide Probes 010606 plant biology & botany Real-time PCR |
| Zdroj: | Fungal Biology Fungal Biology, 2013, 117 (6), pp.389-398. ⟨10.1016/j.funbio.2013.04.001⟩ |
| ISSN: | 1878-6146 |
| DOI: | 10.1016/j.funbio.2013.04.001⟩ |
| Popis: | International audience; Melampsora medusae (Mm), one of the causal agents of poplar rust, is classified as an A2 quarantine pest for European Plant Protection Organization (EPPO) and its presence in Europe is strictly controlled. Two formae speciales have been described within Mm, Melampsora medusae f. sp. deltoidae (Mmd), and Melampsora medusae f. sp. tremuloidae (Mmt) on the basis of their pathogenicity on Populus species from the section Aigeiros (e.g. Populus deltoides) or Populus (e.g. Populus tremuloides), respectively. In this study, a real-time polymerase chain reaction (PCR) assay was developed allowing the detection of Mmd, the forma specialis that is economically harmful. A set of primers and hydrolysis probe were designed based on sequence polymorphisms in the large ribosomal RNA subunit (28S). The real-time PCR assay was optimized and performance criteria of the detection method, i.e. sensitivity, specificity, repeatability, reproducibility, and robustness, were assessed. The real-time PCR method was highly specific and sensitive and allowed the detection of one single urediniospore of Mmd in a mixture of 2 mg of urediniospores of other Melampsora species. This test offers improved specificity over currently existing conventional PCR tests and can be used for specific surveys in European nurseries and phytosanitary controls, in order to avoid introduction and spread of this pathogen in Europe. |
| Databáze: | OpenAIRE |
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