Abstract
Powdery mildew poses a serious problem for apple growers, and resistance to the disease is a major objective in breeding programmes for cultivar improvement. As selective pressure allows pathogens to overcome previously reliable resistances, there is a need for the introduction of novel resistance genes into new breeding lines. This investigation is concerned with the identification of the first set of molecular markers linked to the gene for mildew resistance, Pl-d, from the accession ‘D12’. As no prior information on the map position or markers for Pl-d were available, a bulked-segregant approach was used to test 49 microsatellite primers, 176 amplified fragment length polymorphism (AFLP) primers and 80 random amplified polymorphic DNA (RAPD) primers in a progeny segregating for Pl-d resistance, ‘Fiesta’ (susceptible) × A871-14 (‘Worcester Pearmain’ × ‘D12’). The segregations of the markers identified in the resistant and susceptible bulks were scored in the progeny, then the recombination fractions between Pl-d and the most tightly linked markers were calculated and a map prepared. Three AFLP, one RAPD and two microsatellite markers were identified. One AFLP was developed into a sequence-characterised amplified region marker, while the microsatellites CH03C02 and CH01D03 were flanking markers, 7 and 11 recombination units, respectively, from Pl-d. Two more distant microsatellites on the same linkage group, CH01D09 and CH01G12, confirmed the orientation of the markers on the linkage group. These microsatellites place Pl-d on the bottom of linkage group 12 in published apple maps, a region where a number of other disease resistance genes have been identified.
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Acknowledgements
We thank Bobbie Maxted and Katherine Phillips for their assistance in the glasshouse and field work, Emily Buck for work on the AFLPs, Raul De La Rosa for advice on RAPDs, Robert Liebhard,Cesare Gessler, Bernard Koller and Eric van der Weg for information about the microsatellites and Kenneth Tobutt for valuable discussion and critical reading of the manuscript. This work was part of a Shared-Cost project funded by the European Union (DG VI, FAIR5 CT97-3898) with matching funding from DEFRA. The content of the publication is the sole responsibility of its authors and in no way represents the views of the European Commission or its services.
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James, C.M., Clarke, J.B. & Evans, K.M. Identification of molecular markers linked to the mildew resistance gene Pl-d in apple. Theor Appl Genet 110, 175–181 (2004). https://doi.org/10.1007/s00122-004-1836-0
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DOI: https://doi.org/10.1007/s00122-004-1836-0