Anchoring linkage groups of theRosa genetic map to physical chromosomes with tyramide-FISH and EST-SNP markers

Ilia Kirov, Katrijn Van Laere, Jan De Riek, Ellen De Keyser, Nadine Van Roy, ludmila Khrustaleva

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    Abstract

    In order to anchor Rosa linkage groups to physical chromosomes, a combination of the Tyramide-FISH technology and the modern molecular marker system based on High Resolution Melting (HRM) is an efficient approach. Although, Tyramide-FISH is a very promising technique for the visualization of short DNA probes, it is very challenging for plant species with small chromosomes such as Rosa. In this study, we successfully applied the Tyramide-FISH technique for Rosa and compared different detection systems. An indirect detection system exploiting biotinylated tyramides was shown to be the most suitable technique for reliable signal detection. Three gene fragments with a size of 1100 pb–1700 bp (Phenylalanine Ammonia Lyase, Pyrroline-5-Carboxylate Synthase and Orcinol O-Methyl Transferase) have been physically mapped on chromosomes 7, 4 and 1, respectively, of Rosa wichurana. The signal frequency was between 25% and 40%. HRM markers of these 3 gene fragments were used to include the gene fragments on the existing genetic linkage map of Rosa wichurana. As a result, three linkage groups could be anchored to their physical chromosomes. The information was used to check for synteny between the Rosa chromosomes and Fragaria.
    Original languageEnglish
    JournalPloS one
    Volume9
    Issue number4
    Pages (from-to)e95793
    Number of pages9
    ISSN1932-6203
    DOIs
    Publication statusPublished - 2014

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