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A DNA marker assay based on high-resolution melting curve analysis for distinguishing species of the Festuca - Lolium complex

Birrer, Martina ; Kölliker, Roland ; Manzanares, Chloe ; Asp, Torben ; Studer, Bruno

In: Molecular Breeding, 2014, vol. 34, no. 2, p. 421-429

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    Titre
    • A DNA marker assay based on high-resolution melting curve analysis for distinguishing species of the Festuca - Lolium complex
    Auteur
    • Birrer, Martina. Forage Crop Genetics, Institute of Agricultural Sciences, ETH Zurich, Universitaetstrasse 2, 8092, Zurich, Switzerland
    • Kölliker, Roland. Institute for Sustainability Sciences, ISS, Agroscope, Reckenholzstrasse 196, 8046, Zurich, Switzerland
    • Manzanares, Chloe. Forage Crop Genetics, Institute of Agricultural Sciences, ETH Zurich, Universitaetstrasse 2, 8092, Zurich, Switzerland
    • Asp, Torben. Department of Molecular Biology and Genetics, Faculty of Science and Technology, Aarhus University, Forsøgsvej 1, 4200, Slagelse, Denmark
    • Studer, Bruno. Forage Crop Genetics, Institute of Agricultural Sciences, ETH Zurich, Universitaetstrasse 2, 8092, Zurich, Switzerland
    Type de document
    • Postprint
    Identifiant OAI-PMH
    • oai:doc.rero.ch:324760
    Summary
    The grass breeding industry is interested in a fast and cheap method of identifying contamination in seeds of Italian and perennial ryegrass (Lolium perenne L. and L. multiflorum Lam., respectively). This study shows that high-resolution melting curve analysis in combination with an unlabelled probe assay is an effective method of detecting single nucleotide polymorphisms (SNPs) in diverse Italian and perennial ryegrass backgrounds. This method proved efficient in differentiating ryegrass species and reducing the effect of additional DNA sequence polymorphisms close to the target SNP on the melting curve profiles. For the identification of contamination in Italian and perennial ryegrass seed production, high-resolution melting curve analysis shows great potential, as it is a single closed-tube PCR reaction with an easy workflow, providing results in <2h after DNA extraction.