Advances in flow cytogenetics of barley

Suchánková, Pavla

Flow cytogenetics is an attractive tool that facilitates genome mapping. Flow cytometric analysis permits classification of isolated chromosomes according to their DNA content As thousands of chromosomes are analysed in each sample, quantitative detection of structural and numerical chromosome changes is possible. Any chromosome that can be discriminated can also be sorted in large quantities and high purities for subsequent molecular analysis. Unfortunately, only the smallest chromosome 5(1H) can be discriminated and sorted in barley. This is due to similarity in relative DNA content among the chromosomes 1 - 4,6 and 7 (2H - 7H). Cytogenetic stocks facilitate sorting of those translocation chromosomes that differ in DNA content Even if the sorted translocation chromosomes were found useful for mapping DNA sequences to subchromosomal regions, their use remained limited. Here we describe a novel approach that facilitates sorting of any of the 14 barley chromosome arms. The strategy is based on using wheat-barley addition lines carrying barley chromosome arms (telocentric chromosomes). Furthermore, we will be able to sort subarm chromosomal segments that are generated in the wheat-barley chromosome addition lines by the gametocidal system. These advances make flow cytogenetics an attractive tool that can simplify barley genome mapping. A possibility to purify large quantities of individual chromosome arms and their segments opens avenues for targeted isolation of low-copy "genie" sequences, preparation of specific probes for screening EST arrays, HAPPY mapping, and construction of chromosome arm-specific BAG libraries.

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