Chromosome sorting in tetraploid wheat and its potential for genome analysis

Marie Kubaláková, Pavlína Kovářová, Pavla Suchánková, Jarmila Číhalíková, Jan Bartoš, Sergio Lucretti, Nobuyoshi Watanabe, Shahryar F. Kianian, Jaroslav Doležel

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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 flowcytogenetics for use in wheat genomics and opens the possibility of sequencing the genome of this important crop one chromosome arm at a time. Copyright © 2005 by the Genetics Society of America.
Original languageEnglish
Pages (from-to)823 - 829
Number of pages7
JournalGenetics
Volume170
Issue number2
DOIs
Publication statusPublished - Jun 2005
Externally publishedYes

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All Science Journal Classification (ASJC) codes

  • Genetics

Cite this

Kubaláková, M., Kovářová, P., Suchánková, P., Číhalíková, J., Bartoš, J., Lucretti, S., ... Doležel, J. (2005). Chromosome sorting in tetraploid wheat and its potential for genome analysis. Genetics, 170(2), 823 - 829. https://doi.org/10.1534/genetics.104.039180