Abstract
BackgroundRosaceae is a family containing many economically important fruit and ornamental species. Although fluorescence in situ hybridization (FISH)-based physical mapping of plant genomes is a valuable tool for map-based cloning, comparative genomics and evolutionary studies, no studies using high resolution physical mapping have been performed in this family. Previously we proved that physical mapping of single-copy genes as small as 1.1 kb is possible on mitotic metaphase chromosomes of Rosa wichurana using Tyramide-FISH. In this study we aimed to further improve the physical map of Rosa wichurana by applying high resolution FISH to pachytene chromosomes.ResultsUsing high resolution Tyramide-FISH and multicolor Tyramide-FISH, 7 genes (1.7–3 kb) were successfully mapped on pachytene chromosomes 4 and 7 of Rosa wichurana. Additionally, by using multicolor Tyramide-FISH three closely located genes were simultaneously visualized on chromosome 7. A detailed map of heterochromatine/euchromatine patterns of chromosome 4 and 7 was developed with indication of the physical position of these 7 genes. Comparison of the gene order between Rosa wichurana and Fragaria vesca revealed a poor collinearity for chromosome 7, but a perfect collinearity for chromosome 4.ConclusionsHigh resolution physical mapping of short probes on pachytene chromosomes of Rosa wichurana was successfully performed for the first time. Application of Tyramide-FISH on pachytene chromosomes allowed the mapping resolution to be increased up to 20 times compared to mitotic metaphase chromosomes. High resolution Tyramide-FISH and multicolor Tyramide-FISH might become useful tools for further physical mapping of single-copy genes and for the integration of physical and genetic maps of Rosa wichurana and other members of the Rosaceae.Electronic supplementary materialThe online version of this article (doi:10.1186/s12863-015-0233-9) contains supplementary material, which is available to authorized users.
Highlights
Rosaceae is a family containing many economically important fruit and ornamental species
DAPI staining of R wichurana pachytene chromosomes did not reveal strong heterochromatin blocks that can be used as cytogenetic markers
The SteamDrop protocol was modified by adding a drop of 60 % acetic acid after the second steam application and before drying the cells
Summary
Rosaceae is a family containing many economically important fruit and ornamental species. In this study we aimed to further improve the physical map of Rosa wichurana by applying high resolution FISH to pachytene chromosomes. Rosa is a genus of the Rosaceae family consisting of approximately 90 genera and approximately 3000 species. Many of these are economically important such as Malus, Prunus, Pyrus, Fragaria, Rubus, Sorbus, Cotoneaster and Crataegus [1,2,3,4,5]. Basic chromosome number is x = 7 [1], but Genetic maps have been widely used for comparative genomic and genome organization studies [12, 13].
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