Microsatellite markers for direct genotyping of the crayfish plague pathogen Aphanomyces astaci (Oomycetes) from infected host tissues
| Autor: | Joa Mangombi, Lenka Filipová, Trude Vrålstad, Svetlana Rezinciuc, Carine Delaunay, Eva Kozubíková-Balcarová, Adam Petrusek, Mišel Jelić, Satu Viljamaa-Dirks, Javier Diéguez-Uribeondo, Frédéric Grandjean, Daniel Guyonnet |
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| Přispěvatelé: | Ecologie, Evolution, Symbiose (EES), Ecologie et biologie des interactions (EBI), Université de Poitiers-Centre National de la Recherche Scientifique (CNRS)-Université de Poitiers-Centre National de la Recherche Scientifique (CNRS), Departamento de Micologia, Université de Madrid, Department of Ecology, Charles University [Prague] (CU) |
| Rok vydání: | 2014 |
| Předmět: |
Crayfish plague
Genotype Astacoidea Aphanomyces Microbiology Animals Axenic Genotyping Animals Aphanomyces/classification/*genetics/isolation & purification Astacoidea/*parasitology Europe Genetic Variation Genotype Microsatellite Repeats/*genetics Random Amplified Polymorphic DNA Technique Genetics Oomycete crayfish plague Aphanomyces astaci invasive crayfish crayfish mass mortalities microsatellites genotyping General Veterinary biology musculoskeletal neural and ocular physiology European Crayfish Genetic Variation General Medicine biology.organism_classification Crayfish Random Amplified Polymorphic DNA Technique 3. Good health Europe nervous system Microsatellite [SDE.BE]Environmental Sciences/Biodiversity and Ecology Microsatellite Repeats |
| Zdroj: | Veterinary Microbiology Veterinary Microbiology, Elsevier, 2014, 3-4, pp.317-324. ⟨10.1016/j.vetmic.2014.02.020⟩ |
| ISSN: | 0378-1135 |
| DOI: | 10.1016/j.vetmic.2014.02.020 |
| Popis: | International audience; Aphanomyces astaci is an invasive pathogenic oomycete responsible for the crayfish plague, a disease that has devastated European freshwater crayfish. So far, five genotype groups of this pathogen have been identified by applying random amplified polymorphic DNA analysis on axenic cultures. To allow genotyping of A. astaci in host tissue samples, we have developed co-dominant microsatellite markers for this pathogen, tested them on pure cultures of all genotype groups, and subsequently evaluated their use on tissues of (1) natural A. astaci carriers, i.e., North American crayfish species, and (2) A. astaci-infected indigenous European species from crayfish plague outbreaks. Out of over 200 potential loci containing simple sequence repeat (SSR) motifs identified by 454 pyrosequencing of SSR-enriched library, we tested 25 loci with highest number of repeats, and finally selected nine that allow unambiguous separation of all known RAPD-defined genotype groups of A. astaci from axenic cultures. Using these markers, we were able to characterize A. astaci strains from DNA isolates from infected crayfish tissues when crayfish had a moderate to high agent level according to quantitative PCR analyses. The results support the hypothesis that different North American crayfish hosts carry different genotype groups of the pathogen, and confirm that multiple genotype groups, including the one originally introduced to Europe in the 19th century, cause crayfish plague outbreaks in Central Europe. So far undocumented A. astaci genotype seems to have caused one of the analysed outbreaks from the Czech Republic. The newly developed culture-independent approach allowing direct genotyping of this pathogen in both axenic cultures and mixed genome samples opens new possibilities in studies of crayfish plague pathogen distribution, diversity and epidemiology. |
| Databáze: | OpenAIRE |
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