Advanced resources for plant genomics: a BAC library specific for the short arm of wheat chromosome 1B

Authors

  • Jaroslav Janda,

    1. Laboratory of Molecular Cytogenetics and Cytometry, Institute of Experimental Botany, Sokolovská 6, CZ-77200 Olomouc, Czech Republic,
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  • Jan Šafář,

    1. Laboratory of Molecular Cytogenetics and Cytometry, Institute of Experimental Botany, Sokolovská 6, CZ-77200 Olomouc, Czech Republic,
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  • Marie Kubaláková,

    1. Laboratory of Molecular Cytogenetics and Cytometry, Institute of Experimental Botany, Sokolovská 6, CZ-77200 Olomouc, Czech Republic,
    2. Department of Cell Biology and Genetics, Palacký University, Šlechtitelů 11, CZ-78371 Olomouc, Czech Republic,
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  • Jan Bartoš,

    1. Laboratory of Molecular Cytogenetics and Cytometry, Institute of Experimental Botany, Sokolovská 6, CZ-77200 Olomouc, Czech Republic,
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  • Pavlína Kovářová,

    1. Laboratory of Molecular Cytogenetics and Cytometry, Institute of Experimental Botany, Sokolovská 6, CZ-77200 Olomouc, Czech Republic,
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  • Pavla Suchánková,

    1. Laboratory of Molecular Cytogenetics and Cytometry, Institute of Experimental Botany, Sokolovská 6, CZ-77200 Olomouc, Czech Republic,
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  • Stephanie Pateyron,

    1. Organization and Evolution of Plant Genomes, Unité de Recherches en Génomique Végétale (INRA-URGV), 2 rue Gaston Crémieux, CP 5708, F-91057 Évry Cedex, France,
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  • Jarmila Číhalíková,

    1. Laboratory of Molecular Cytogenetics and Cytometry, Institute of Experimental Botany, Sokolovská 6, CZ-77200 Olomouc, Czech Republic,
    2. Department of Cell Biology and Genetics, Palacký University, Šlechtitelů 11, CZ-78371 Olomouc, Czech Republic,
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  • Pierre Sourdille,

    1. Génétique Moléculaire des Céréales, UMR INRA-UBP, Domaine de Crouelle, 234 Avenue du Brézet, F-63039 Clermont-Ferrand Cedex 2, France, and
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  • Hana Šimková,

    1. Laboratory of Molecular Cytogenetics and Cytometry, Institute of Experimental Botany, Sokolovská 6, CZ-77200 Olomouc, Czech Republic,
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  • Patricia Faivre-Rampant,

    1. Organization and Evolution of Plant Genomes, Unité de Recherches en Génomique Végétale (INRA-URGV), 2 rue Gaston Crémieux, CP 5708, F-91057 Évry Cedex, France,
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  • Eva Hřibová,

    1. Laboratory of Molecular Cytogenetics and Cytometry, Institute of Experimental Botany, Sokolovská 6, CZ-77200 Olomouc, Czech Republic,
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  • Michel Bernard,

    1. Génétique Moléculaire des Céréales, UMR INRA-UBP, Domaine de Crouelle, 234 Avenue du Brézet, F-63039 Clermont-Ferrand Cedex 2, France, and
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  • Adam Lukaszewski,

    1. Department of Botany and Plant Sciences, University of California, Riverside, CA 92521, USA
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  • Jaroslav Doležel,

    Corresponding author
    1. Laboratory of Molecular Cytogenetics and Cytometry, Institute of Experimental Botany, Sokolovská 6, CZ-77200 Olomouc, Czech Republic,
    2. Department of Cell Biology and Genetics, Palacký University, Šlechtitelů 11, CZ-78371 Olomouc, Czech Republic,
      (fax + 420 585 205 853; e-mail dolezel@ueb.cas.cz).
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  • Boulos Chalhoub

    1. Organization and Evolution of Plant Genomes, Unité de Recherches en Génomique Végétale (INRA-URGV), 2 rue Gaston Crémieux, CP 5708, F-91057 Évry Cedex, France,
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(fax + 420 585 205 853; e-mail dolezel@ueb.cas.cz).

Summary

Common wheat (Triticum aestivum L., 2n = 6x = 42) is a polyploid species possessing one of the largest genomes among the cultivated crops (1C is approximately 17 000 Mb). The presence of three homoeologous genomes (A, B and D), and the prevalence of repetitive DNA make sequencing the wheat genome a daunting task. We have developed a novel ‘chromosome arm-based’ strategy for wheat genome sequencing to simplify this task; this relies on sub-genomic libraries of large DNA inserts. In this paper, we used a di-telosomic line of wheat to isolate six million copies of the short arm of chromosome 1B (1BS) by flow sorting. Chromosomal DNA was partially digested with HindIII and used to construct an arm-specific BAC library. The library consists of 65 280 clones with an average insert size of 82 kb. Almost half of the library (45%) has inserts larger than 100 kb, while 18% of the inserts range in size between 75 and 100 kb, and 37% are shorter than 75 kb. We estimated the chromosome arm coverage to be 14.5-fold, giving a 99.9% probability of identifying a clone corresponding to any sequence on the short arm of 1B. Each chromosome arm in wheat can be flow sorted from an appropriate cytogenetic stock, and we envisage that the availability of chromosome arm-specific BAC resources in wheat will greatly facilitate the development of ready-to-sequence physical maps and map-based gene cloning.

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