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

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 D...

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Bibliographic Details
Published in:PloS one 2014-04, Vol.9 (4), p.e95793-e95793
Main Authors: Kirov, Ilya, Van Laere, Katrijn, De Riek, Jan, De Keyser, Ellen, Van Roy, Nadine, Khrustaleva, Ludmila
Format: Article
Language:English
Subjects:
Ammonia
Anchoring
Biology and Life Sciences
Biotechnology
Chromosome Mapping
Chromosomes
Chromosomes, Plant
Deoxyribonucleic acid
DNA
DNA probes
Expressed Sequence Tags
Fish
Fisheries
Fluorescence in situ hybridization
Fragaria
Fragaria vesca
Fragmentation
Fragments
Gene expression
Genetic aspects
Genetic Linkage
Genetic Markers
Genetics
Genomes
Hybridization
In Situ Hybridization, Fluorescence - methods
Linkage (Genetics)
Markers
Molecular biology
Phenylalanine
Physiological aspects
Plant sciences
Polymorphism, Single Nucleotide
Research and Analysis Methods
Rosa - genetics
Roses
Signal detection
Single-nucleotide polymorphism
Synteny
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Summary: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.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0095793