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Construction and characterization of a half million clone BAC library of durum wheat (Triticum turgidum ssp. durum)

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Abstract.

Durum wheat (Triticum turgidum ssp. durum, 2n = 4x = 28, genomes AB) is an economically important cereal used as the raw material to make pasta and semolina. In this paper we present the construction and characterization of a bacterial artificial chromosome (BAC) library of tetraploid durum wheat cv. Langdon. This variety was selected because of the availability of substitution lines that facilitate the assignment of BACs to the A and B genome. The selected Langdon line has a 30-cM segment of chromosome 6BS from T. turgidum ssp. dicoccoides carrying a gene for high grain protein content, the target of a positional cloning effort in our laboratory. A total of 516,096 clones were organized in 1,344 384-well plates and blotted on 28 high-density filters. Ninety-eight percent of these clones had wheat DNA inserts (0.3% chloroplast DNA, 1.4% empty clones and 0.3% empty wells). The average insert size of 500 randomly selected BAC clones was 131 kb, resulting in a coverage of 5.1-fold genome equivalents for each of the two genomes, and a 99.4% probability of recovering any gene from each of the two genomes of durum wheat. Six known copy-number probes were used to validate this theoretical coverage and gave an estimated coverage of 5.8-fold genome equivalents. Screening of the library with 11 probes related to grain storage proteins and starch biosynthesis showed that the library contains several clones for each of these genes, confirming the value of the library in characterizing the organization of these important gene families. In addition, characterization of fingerprints from colinear BACs from the A and B genomes showed a large differentiation between the A and B genomes. This library will be a useful tool for evolutionary studies in one of the best characterized polyploid systems and a source of valuable genes for wheat. Clones and high-density filters can be requested at http://agronomy.ucdavis.edu/Dubcovsky/BAC-library/BAC_Langdon.htm

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Acknowledgements.

This research was supported by Research Grant No. US-3224-01R from BARD (The United States -Israel Binational Agricultural Research and Development Fund), by INRA-France and by USDA-Albany. The authors thank Dr R. N. Chibbar for the screening of the Langdon BAC library with the genes involved in starch biosynthesis.

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Author notes

  1. A. Cenci

    Present address: Dip. Biologia e Chimica Agroforestale e Ambientale, Università degli Studi di Bari, Via G. Amendola 165/a, 70126 Bari, Italy,

  2. N. Chantret

    Present address: CIRAD-AMIS-Biotrop, av. Agropolis, TA 40/30, 34398 Montpellier Cedex 5, France,

  3. T. Fahima & A. Distelfeld

    Present address: The Institute of Evolution, University of Haifa, Mt. Carmel, Haifa. Israel,

Authors and Affiliations

  1. Department of Agronomy and Range Science, University of California, One Shields Avenue, Davis, CA 95616-8515, USA

    A. Cenci, N. Chantret, T. Fahima, A. Distelfeld & J. Dubcovsky

  2. USDA Western Reg. Research Center, 800 Buchanan St, Albany, CA 94710, USA

    X. Kong, Y. Gu & O. D. Anderson

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  1. A. Cenci
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  2. N. Chantret
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  3. X. Kong
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Correspondence to J. Dubcovsky.

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Communicated by P. Langridge

The first two authors contributed equally to the investigation

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Cenci, A., Chantret, N., Kong, X. et al. Construction and characterization of a half million clone BAC library of durum wheat (Triticum turgidum ssp. durum). Theor Appl Genet 107, 931–939 (2003). https://doi.org/10.1007/s00122-003-1331-z

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  • DOI: https://doi.org/10.1007/s00122-003-1331-z

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