Anchoring linkage groups of the Rosa genetic map to physical chromosomes with tyramide-FISH and EST-SNP markers.
TL;DR: An indirect detection system exploiting biotinylated tyramides was shown to be the most suitable technique for reliable signal detection and compared different detection systems.
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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.
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Citations
Fluorescence in situ hybridization in plants: recent developments and future applications.
TL;DR: The most significant recent advance of FISH is the development and application of probes based on synthetic oligonucleotides (oligos) which can be computationally identified, synthesized in parallel, and fluorescently labeled.
171
High-Resolution DNA Melting Analysis in Plant Research
TL;DR: Use of HRM has expanded considerably in the past few years as the method was successfully applied for high-throughput genotyping, mapping genes, testing food products and seeds, and other areas of plant research.
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High-density SNP-based genetic maps for the parents of an outcrossed and a selfed tetraploid garden rose cross, inferred from admixed progeny using the 68k rose SNP array.
M. Vukosavljev,Paul Arens,Roeland E. Voorrips,Wendy van’t Westende,G.D. Esselink,Peter M. Bourke,Peter Cox,W. Eric van de Weg,Richard G. F. Visser,Chris Maliepaard,Marinus J. M. Smulders +10 more
TL;DR: A strategy to separate progeny into putative populations, even while one of the parents was unknown, was developed, using principle component analysis on pairwise genetic distances based on sets of selected SNP markers that were homozygous, and therefore uninformative for one parent.
High-resolution tyramide-FISH mapping of markers tightly linked to the male-fertility restoration (Ms) locus of onion
TL;DR: The results indicate that map-based cloning of Ms will likely be difficult due to reduced recombination near this gene, and explains why several labs have identified molecular markers tightly linked to the Ms locus after screening relatively few DNA clones or primers and segregating progenies.
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Mapping of single-copy genes by TSA-FISH in the codling moth, Cydia pomonella.
TL;DR: The results suggest that this technique can be combined with BAC-FISH and in the future used for physical localization of transgene cassettes on chromosomes of transgenic lines in the codling moth or other lepidopteran species.
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