1. A rapid genotyping method for an obligate fungal pathogen, Puccinia striiformis f.sp. tritici, based on DNA extraction from infected leaf and Multiplex PCR genotyping
- Author
-
Jérôme Enjalbert, Sajid Ali, Angélique Gautier, Marc Leconte, Claude de Vallavieille-Pope, BIOlogie et GEstion des Risques en agriculture (BIOGER), AgroParisTech-Institut National de la Recherche Agronomique (INRA), Génétique Quantitative et Evolution - Le Moulon (Génétique Végétale) (GQE-Le Moulon), Centre National de la Recherche Scientifique (CNRS)-AgroParisTech-Université Paris-Sud - Paris 11 (UP11)-Institut National de la Recherche Agronomique (INRA), and Institut National de la Recherche Agronomique (INRA)-Université Paris-Sud - Paris 11 (UP11)-AgroParisTech-Centre National de la Recherche Scientifique (CNRS)
- Subjects
0106 biological sciences ,[SDV.SA]Life Sciences [q-bio]/Agricultural sciences ,genotyping method ,multiplex pcr genotyping ,fungal pathogen ,genotype ,microsatellitte ,rouille jaune du blé ,Puccinia striiformis ,diversité génétique ,lcsh:Medicine ,Biology ,01 natural sciences ,Rust ,General Biochemistry, Genetics and Molecular Biology ,03 medical and health sciences ,Genotype ,Multiplex polymerase chain reaction ,Technical Note ,lcsh:Science (General) ,Pathogen ,Genotyping ,lcsh:QH301-705.5 ,030304 developmental biology ,2. Zero hunger ,Genetics ,Medicine(all) ,0303 health sciences ,Molecular epidemiology ,Obligate ,Biochemistry, Genetics and Molecular Biology(all) ,lcsh:R ,food and beverages ,General Medicine ,DNA extraction ,lcsh:Biology (General) ,010606 plant biology & botany ,lcsh:Q1-390 - Abstract
Background Puccinia striiformis f.sp. tritici (PST), an obligate fungal pathogen causing wheat yellow/stripe rust, a serious disease, has been used to understand the evolution of crop pathogen using molecular markers. However, numerous questions regarding its evolutionary history and recent migration routes still remains to be addressed, which need the genotyping of a large number of isolates, a process that is limited by both DNA extraction and genotyping methods. To address the two issues, we developed here a method for direct DNA extraction from infected leaves combined with optimized SSR multiplexing. Findings We report here an efficient protocol for direct fungal DNA extraction from infected leaves, avoiding the costly and time consuming step of spore multiplication. The genotyping strategy we propose, amplified a total of 20 SSRs in three Multiplex PCR reactions, which were highly polymorphic and were able to differentiate different PST populations with high efficiency and accuracy. Conclusion These two developments enabled a genotyping strategy that could contribute to the development of molecular epidemiology of yellow rust disease, both at a regional or worldwide scale.
- Published
- 2011
- Full Text
- View/download PDF