title: Chromosome sorting in tetraploid wheat and its potential for genome analysis.
abstract: This study evaluates the potential of flow cytometry for chromosome sorting in durum wheat (Triticum turgidum Desf. var. durum, 2n = 4x = 28). Histograms of fluorescence intensity (flow karyotypes) obtained after the analysis of DAPI-stained chromosomes consisted of three peaks. Of these, one represented chromosome 3B, a small peak corresponded to chromosomes 1A and 6A, and a large peak represented the remaining 11 chromosomes. Chromosomes sorted onto microscope slides were identified after fluorescence in situ hybridization (FISH) with probes for GAA microsatellite, pSc119.2, and Afa repeats. Genomic distribution of these sequences was determined for the first time in durum wheat and a molecular karyotype has been developed for this crop. Flow karyotyping in double-ditelosomic lines of durum wheat revealed that the lines facilitated sorting of any arm of the wheat A- and B-genome chromosomes. Compared to hexaploid wheat, flow karyotype of durum wheat is less complex. This property results in better discrimination of telosomes and high purities in sorted fractions, ranging from 90 to 98%. We have demonstrated that large insert libraries can be created from DNA purified using flow cytometry. This study considerably expands the potential of flow cytogenetics for use in wheat genomics and opens the possibility of sequencing the genome of this important crop one chromosome arm at a time.
authors.affiliation: Laboratory of Molecular Cytogenetics and Cytometry, Institute of Experimental Botany, Olomouc, Czech Republic.
authors.forename: Marie
authors.initials: M
authors.lastname: Kubaláková
authors.forename: Pavlína
authors.initials: P
authors.lastname: Kovárová
authors.forename: Pavla
authors.initials: P
authors.lastname: Suchánková
authors.forename: Jarmila
authors.initials: J
authors.lastname: Cíhalíková
authors.forename: Jan
authors.initials: J
authors.lastname: Bartos
authors.forename: Sergio
authors.initials: S
authors.lastname: Lucretti
authors.forename: Nobuyoshi
authors.initials: N
authors.lastname: Watanabe
authors.forename: Shahryar F
authors.initials: SF
authors.lastname: Kianian
authors.forename: Jaroslav
authors.initials: J
authors.lastname: Dolezel
mesh_terms: D002453:Cell Cycle; D002469:Cell Separation; D002874:Chromosome Mapping; D002875:Chromosomes / Q000648:ultrastructure; D032461:Chromosomes, Plant; D004247:DNA / Q000235:genetics; D018744:DNA, Plant; D005434:Flow Cytometry; D005821:Genetic Techniques; D016678:Genome; D018745:Genome, Plant; D017404:In Situ Hybridization, Fluorescence; D007621:Karyotyping; D008677:Metaphase; D018895:Microsatellite Repeats; D008957:Models, Genetic; D020161:Physical Chromosome Mapping; D011003:Ploidies; D014908:Triticum / Q000235:genetics*
chemical_list: D018744:DNA, Plant; D004247:DNA