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Dissection of the nuclear genome of barley by chromosome flow sorting

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Abstract

Isolation of mitotic chromosomes using flow cytometry is an attractive way to dissect nuclear genomes into their individual chromosomal components or portions of them. This approach is especially useful in plants with complex genomes, where it offers a targeted and hence economical approach to genome analysis and gene cloning. In several plant species, DNA of flow-sorted chromosomes has been used for isolation of molecular markers from specific genome regions, for physical mapping using polymerase chain reaction (PCR) and fluorescence in situ hybridization (FISH), for integration of genetic and physical maps and for construction of chromosome-specific DNA libraries, including those cloned in bacterial artificial chromosome vectors. Until now, chromosome analysis and sorting using flow cytometry (flow cytogenetics) has found little application in barley (2n = 14, 1C ∼ 5,100 Mbp) because of the impossibility of discriminating and sorting individual chromosomes, except for the smallest chromosome 1H and some translocation chromosomes with DNA content significantly different from the remaining chromosomes. In this work, we demonstrate that wheat–barley ditelosomic addition lines can be used to sort any arm of barley chromosomes 2H–7H. Thus, the barley genome can be dissected into fractions representing only about 6–12% of the total genome. This advance makes the flow cytogenetics an attractive tool, which may greatly facilitate genome analysis and gene cloning in barley.

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Acknowledgments

We are grateful to our colleagues Jitka Weiserová, Radka Tušková, Michaela Libosvárová and Romana Nováková for excellent technical assistance, and Dr Hana Šimková for improvement of this manuscript. This work was supported by research grants from the Czech Science Foundation (grant award 521/05/H013), Ministry of Education, Youth and Sports of the Czech Republic (grant awards ME884 and LC06004), and was performed within a framework of a collaborative agreement No. 14 between the Academy of Sciences of the Czech Republic and the Hungarian Academy of Sciences. We thank two anonymous reviewers for valuable suggestions.

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Authors and Affiliations

  1. Laboratory of Molecular Cytogenetics and Cytometry, Institute of Experimental Botany, Sokolovská 6, 77200, Olomouc, Czech Republic

    Pavla Suchánková, Marie Kubaláková, Pavlína Kovářová, Jan Bartoš, Jarmila Číhalíková & Jaroslav Doležel

  2. Department of Cell Biology and Genetics, Palacký University, Olomouc, Czech Republic

    Marie Kubaláková, Jarmila Číhalíková & Jaroslav Doležel

  3. Agricultural Research Institute of the Hungarian Academy of Sciences, Martonvásár, Hungary

    Márta Molnár-Láng

  4. Laboratory of Plant Genetics, Graduate School of Agriculture, Kyoto University, Kyoto, Japan

    Takashi R. Endo

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  1. Pavla Suchánková
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  2. Marie Kubaláková
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Correspondence to Jaroslav Doležel.

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Communicated by J. W. Snape

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Suchánková, P., Kubaláková, M., Kovářová, P. et al. Dissection of the nuclear genome of barley by chromosome flow sorting. Theor Appl Genet 113, 651–659 (2006). https://doi.org/10.1007/s00122-006-0329-8

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