Dissection of the nuclear genome of barley by chromosome flow sorting

Pavla Suchánková, Marie Kubaláková, Pavlína Kovářová, Jan Bartoš, Jarmila Číhalíková, Márta Molnár-Láng, Takashi R. Endo, Jaroslav Doležel

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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.

Original languageEnglish
Pages (from-to)651-659
Number of pages9
JournalTheoretical and Applied Genetics
Volume113
Issue number4
DOIs
Publication statusPublished - Aug 1 2006

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ASJC Scopus subject areas

  • Biotechnology
  • Agronomy and Crop Science
  • Genetics

Cite this

Suchánková, P., Kubaláková, M., Kovářová, P., Bartoš, J., Číhalíková, J., Molnár-Láng, M., Endo, T. R., & Doležel, J. (2006). Dissection of the nuclear genome of barley by chromosome flow sorting. Theoretical and Applied Genetics, 113(4), 651-659. https://doi.org/10.1007/s00122-006-0329-8