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The use of bulk segregant analysis to identify a RAPD marker linked to leaf rust resistance in barley
Authors:D M E Poulsen  R J Henry  R P Johnston  J A G Irwin  R G Rees
Institution:(1) Queensland Department of Primary Industries, Hermitage Research Station, 4370 Warwick, Australia;(2) Co-operative Research Centre for Tropical Plant Pathology, University of Queensland, 4072, Australia;(3) Queensland Agricultural Biotechnology Centre, University of Queensland, 4072, Australia;(4) Queensland Wheat Research Institute, 4350 Toowoomba, Australia
Abstract:An F2 population from a cross between barley accession Q21861 and the Australian barley variety lsquoGalleonrsquo was used to develop RAPD markers for resistance to barley leaf rust (Puccinia hordei). Resistant and susceptible DNA bulks were constructed following the classification of F2 plants by leaf rust infection type. Bulked segregant analysis was then used to identify a 2.7-kb marker, designated OU022700 and located approximately 12cM from RphQ, a leaf rust resistance gene in Q21861. The marker was generated by PCR with the oligonucleotide primer OPU-02 (Operon). Infection types of F3 progeny were used to confirm assignment of F2 genotypes. OU022700 was shown, retrospectively, to be useful in the identification of individual F2 plants that had been originally misclassified as having susceptible infection types. Both the RAPD marker and RphQ will be potentially useful in the development of new barley cultivars.
Keywords:Barley  Puccinia hordei  Resistance genes  Bulk segregant analysis  Polymerase chain reaction
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