Wheat in a Global Environment

0100780 - UEB-Q 20043013 RIV NL eng M - Monography Chapter
Vrána, Jan - Šimková, Hana - Kubaláková, Marie - Čihalíková, Jarmila - Lysák, Martin - Doležel, Jaroslav
Sorting of mitotic chromosomes in common wheat (Triticum aestivum L.) using flow cytometry.
[Sorting of mitotic chromosomes in common wheat (Triticum aestivum L.) using flow cytometry.]
Wheat in a Global Environment. Dordrecht: Kluwer, 2001, s. 531-536
R&D Projects: GA AV ČR IBS5038104
Institutional research plan: CEZ:AV0Z5038910
Keywords : chromosome isolation * chromosome sorting * flow cytometry
Subject RIV: EB - Genetics ; Molecular Biology

The aim of this study was to develop an improved procedure for preparation of chromosome suspensions, and to evaluate the potential of flow cytometry for chromosome sorting in wheat. Suspensions of intact chromosomes were prepared by mechanical homogenization of synchronized root tips after mild fixation with formaldehyde. Histograms of relative fluorescence intensity (flow karyotypes) obtained after the analysis of DAPI-stained chromosomes were characterized and the chromosome content of all peaks on wheat flow karyotype was determined for the first time. Only chromosome 3B could be discriminated on flow karyotypes of wheat lines with standard karyotype. Remaining chromosomes formed three composite peaks and could be sorted only as groups. Chromosome 3B could be sorted at purity >95% as determined by microscopic evaluation of sorted fractions that were labeled using C-PRINS with primers for GAA microsatellites and for Afa repeats, respectively.

Sorting of mitotic chromosomes in common wheat (Triticum aestivum L.) using flow cytometry.
Permanent Link: http://hdl.handle.net/11104/0008269