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The barley Frost resistance-H2 locus

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Abstract

Frost resistance-H2 (Fr-H2) is a major QTL affecting freezing tolerance in barley, yet its molecular basis is still not clearly understood. To gain a better insight into the structural characterization of the locus, a high-resolution linkage map developed from the Nure × Tremois cross was initially implemented to map 13 loci which divided the 0.602 cM total genetic distance into ten recombination segments. A PCR-based screening was then applied to identify positive bacterial artificial chromosome (BAC) clones from two genomic libraries of the reference genotype Morex. Twenty-six overlapping BACs from the integrated physical-genetic map were 454 sequenced. Reads assembled in contigs were subsequently ordered, aligned and manually curated in 42 scaffolds. In a total of 1.47 Mbp, 58 protein-coding sequences were identified, 33 of which classified according to similarity with sequences in public databases. As three complete barley C-repeat Binding Factors (HvCBF) genes were newly identified, the locus contained13 full-length HvCBFs, four Related to AP2 Triticeae (RAPT) genes, and at least five CBF pseudogenes. The final overall assembly of Fr-H2 includes more than 90 % of target region: all genes were identified along the locus, and a general survey of Repetitive Elements obtained. We believe that this gold-standard sequence for the Morex Fr-H2 will be a useful genomic tool for structural and evolutionary comparisons with Fr-H2 in winter-hardy cultivars along with Fr-2 of other Triticeae crops.

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Acknowledgments

We are grateful to J Knibbiche, A Kusserow, and S Koenig for their help in BAC clone handling and Roche/454 sequencing, and to G Malagoli Tagliazucchi and S Bicciato for assistance in bioinformatic analyses. The work was partially funded by the Fondazione Cassa di Risparmio di Modena, Progetto di Ricerca Internazionale FROSTMAP “Physical mapping of the barley Frost resistance-H2 (Fr-H2) locus”, and by the GENOMORE project of the Fondazione Cassa di Risparmio di Carpi (http://www.fondazionecrcarpi.it/).

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  1. Burkhard Steuernagel

    Present address: The Sainsbury Laboratory, John Innes Centre (JIC), Norwich, UK

Authors and Affiliations

  1. Department of Life Sciences, University of Modena and Reggio Emilia, Via Amendola 2, Pad. Besta, 42122, Reggio Emilia, Italy

    Marianna Pasquariello, Enrico Tagliafico, Nicola Pecchioni & Enrico Francia

  2. Center for Genome Research (CGR), University of Modena and Reggio Emilia, Reggio Emilia, Italy

    Marianna Pasquariello, Francesco Gandolfi, Elena Tenedini, Isabella Bernardis, Enrico Tagliafico, Nicola Pecchioni & Enrico Francia

  3. Genomics Research Centre, Consiglio per la ricerca e la sperimentazione in agricoltura (CRA), Fiorenzuola d’Arda, Italy

    Delfina Barabaschi

  4. Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), Gatersleben, Germany

    Axel Himmelbach, Burkhard Steuernagel, Ruvini Ariyadasa & Nils Stein

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  1. Marianna Pasquariello
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  2. Delfina Barabaschi
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Correspondence to Enrico Francia.

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Pasquariello, M., Barabaschi, D., Himmelbach, A. et al. The barley Frost resistance-H2 locus. Funct Integr Genomics 14, 85–100 (2014). https://doi.org/10.1007/s10142-014-0360-9

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