Visualizing chromosome territories and nuclear architecture of large plant genomes using alien introgressions

Perničková, K., Kopecký, D
Plant cytogenetics and cytogenomics: Methods and Protocols 2672: 365-376, 2023

Keywords: Chromosome territory, Interphase nucleus, 3D FISH, Plant hybrid, Confocal microscopy, Imaris
Abstract: Visualization of chromosome territories is a challenging task in plant genomes due to the lack of chromosome-specific probes, especially in species with large genomes. On the other hand, combination of flow sorting, genomic in situ hybridization (GISH), confocal microscopy, and employment of software for 3D modeling enables to visualize and characterize chromosome territories (CT) in interspecific hybrids. Here, we describe the protocol for the analysis of CTs in wheat–rye and wheat–barley hybrids, including amphiploids and introgression forms, where a pair of chromosomes or chromosome arms from one species is introgressed into the genome of another species. In this way, the architecture and dynamics of CTs in various tissues and different stages of cell cycle can be analyzed.
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IEB authors: David Kopecky, Kateřina Perničková