Integration of genetic and physical maps for Rosa wichurana using Tyramide-FISH

Ilya Kirov, Katrijn Van Laere, Ludmila Khrustaleva, Ellen De Keyser, Jan De Riek

    Onderzoeksoutput: Hoofdstuk in Boek/Rapport/CongresprocedureC3: Congres abstract


    The integration of genetic and chromosomal maps is still very challenging for most plant species. Physical maps linked to genetically mapped markers are very valuable for plant breeding, especially when they are integrated with genomic sequence data. The most promising approach to integrate physical and genetic maps is to directly visualize DNA fragments, derived from genetically mapped markers, using FISH (Fluorescent In Situ Hybridization). However, because of the sometimes short length of the DNA fragments, conventional FISH is not always sensitive enough to visualize them. Therefore, Tyramide-FISH, combining the advantage of an enzymatic and a fluorescence-based detection, has great potential for physical mapping of short DNA fragments.
    In previous research a genetic map based on a diploid population R.wichurana x R. ‘Yesterday’ was constructed (Moghaddam et al. 2012). As a proof of concept, we physically map Phenylalanine ammonia-lyase (1700bp), Orcinol O-methyltransferase (1100bp) and Pyrroline-5-carboxylate synthase (1700bp) using optimized Tyramide-FISH for Rosa wichurana. Optimization of the Tyramide FISH protocol was necessary as R. wichurana is a difficult species for cytogenetics because its thiny roots, hard cell wall, low mitotic index, very small chromosomes (size between 0.86 and 1.52 µm) and high polysaccharide content in the tissues. Also EST-SNP markers were designed for these 3 genes and detected by High Resolution Melting; their genetic mapping has been performed. The HRM markers were incorporated into the known genetic maps of Rosa (Moghaddam et al. 2012) and obtained data allowed us to integrate the genetic and physical maps of chromosomes 1, 4 and 7 of Rosa wichurana.
    TitelSixth International Symposium on Rose Research and Cultivation: abstracts
    EditorsThomas Debener
    StatusGepubliceerd - 2013
    Evenement6th International Symposium on Rose Research and Cultivation - Hannover, Duitsland
    Duur: 25-aug-201330-sep-2013


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