caa a22 4500 463159691 CHVBK 20180406164744.0 cr unu---uuuuu 170326e20070301xx s 000 0 eng 10.1007/s10535-007-0009-9 doi (NATIONALLICENCE)springer-10.1007/s10535-007-0009-9 Chromosome analysis and sorting in Vicia sativa using flow cytometry [Elektronische Daten] [P. Kovářová, A. Navrátilová, J. Macas, J. Doležel] Procedures were developed for flow cytometric analysis and sorting of mitotic chromosomes (flow cytogenetics) of common vetch (Vicia sativa L., 2n=12). Suspensions of intact chromosomes were prepared from root tips after cell cycle synchronization, formaldehyde fixation, and mechanical homogenization. On average, 3 × 105 morphologically intact chromosomes could be isolated from 25 root tips. Flow cytometric analysis of DAPI-stained chromosomes resulted in histograms of relative fluorescence intensity (flow karyotypes) containing four peaks, representing particular chromosomes and/or pairs of chromosomes with similar relative DNA content. Peaks I and II were assigned to chromosomes 6 and 5, respectively. These chromosomes could be sorted with a purity exceeding 90 %. The two remaining peaks on the flow karyotype were composite, each of them representing a pair of chromosomes. Chromosomes 1 and 3 were assigned to composite peak III while chromosomes 2 and 4 were assigned to composite peak IV. The chromosomes could be sorted with a purity of 99 % from both composite peaks. Bivariate flow karyotyping after simultaneous staining of chromosomes with DAPI and mithramycin was not found helpful in discriminating additional chromosomes. This study extends the number of legume species for which flow cytogenetics is available and provides a new tool for targeted and effective analysis and mapping of common vetch genome. Institute of Experimental Botany, ASCR, 2007 cell cycle synchronization nationallicence chromosome suspension nationallicence common vetch nationallicence metaphase accumulation nationallicence physical genome mapping nationallicence DAPI : 4′,6-diamino-2-phenylindole nationallicence FISH : fluorescence in situ hybridization nationallicence HU : hydroxyurea nationallicence PRINS : primed in situ DNA labelling nationallicence Kovářová P. Laboratory of Molecular Cytogenetics and Cytometry, Institute of Experimental Botany, Sokolovská 6, CZ-77200, Olomouc, Czech Republic aut Navrátilová A. Institute of Plant Molecular Biology, Branišovská 31, CZ-37005, České Budějovice, Czech Republic aut Macas J. Institute of Plant Molecular Biology, Branišovská 31, CZ-37005, České Budějovice, Czech Republic aut Doležel J. Laboratory of Molecular Cytogenetics and Cytometry, Institute of Experimental Botany, Sokolovská 6, CZ-77200, Olomouc, Czech Republic aut Biologia Plantarum Springer Netherlands 51/1(2007-03-01), 43-48 0006-3134 51:1<43 2007 51 10535 https://doi.org/10.1007/s10535-007-0009-9 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10535-007-0009-9 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Kovářová P. Laboratory of Molecular Cytogenetics and Cytometry, Institute of Experimental Botany, Sokolovská 6, CZ-77200, Olomouc, Czech Republic aut NATIONALLICENCE 700 1- Navrátilová A. Institute of Plant Molecular Biology, Branišovská 31, CZ-37005, České Budějovice, Czech Republic aut NATIONALLICENCE 700 1- Macas J. Institute of Plant Molecular Biology, Branišovská 31, CZ-37005, České Budějovice, Czech Republic aut NATIONALLICENCE 700 1- Doležel J. Laboratory of Molecular Cytogenetics and Cytometry, Institute of Experimental Botany, Sokolovská 6, CZ-77200, Olomouc, Czech Republic aut NATIONALLICENCE 773 0- Biologia Plantarum Springer Netherlands 51/1(2007-03-01), 43-48 0006-3134 51:1<43 2007 51 10535 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer