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The use of high resolution melting (HRM) to map single nucleotide polymorphism markers linked to a covered smut resistance gene in barley

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Using an established genetic map, a single gene conditioning covered smut resistance, Ruh.7H, was mapped to the telomere region of chromosome 7HS in an Alexis/Sloop doubled haploid barley population. The closest marker to Ruh.7H, abg704 was 7.5 cM away. Thirteen loci on the distal end of 7HS with potential to contain single nucleotide polymorphisms (SNPs) were identified by applying a comparative genomics approach using rice sequence data. Of these, one locus produced polymorphic co-dominant bands of different size while two further loci contained SNPs that were identified using the recently developed high resolution melting (HRM) technique. Two of these markers flanked Ruh.7H with the proximal marker located 3.8 cM and the distal marker 2.7 cM away. This is the first report on the application of the HRM technique to SNP detection and to rapid scoring of known cleaved amplified polymorphic sequence (CAPS) markers in plants. This simple, precise post-PCR technique should find widespread use in the fine-mapping of genetic regions of interest in complex cereal and other plant genomes.

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This project was funded by the Grains Research & Development Corporation’s Australian Winter Cereals Molecular Marker Program. We would like to thank Dr Emma Mace and Noel Knight for valuable discussions, Dr Graham Wildermuth and Greg Platz for their advice on phenotyping, and Dr Christine McDonald and Dr Ashley Plank for assistance with statistical analysis. Maria Harris assisted RBM with the covered smut phenotyping.

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Correspondence to Anke Lehmensiek.

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Communicated by F. Ordon.

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Lehmensiek, A., Sutherland, M.W. & McNamara, R.B. The use of high resolution melting (HRM) to map single nucleotide polymorphism markers linked to a covered smut resistance gene in barley. Theor Appl Genet 117, 721–728 (2008).

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