A strategy to develop molecular markers applicable to a wide range of crosses for marker assisted selection in plant breeding: a case study on anthracnose disease resistance in lupin (Lupinus angustifolius L.) |
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Authors: | Huaan Yang Daniel Renshaw Geoff Thomas Bevan Buirchell Mark Sweetingham |
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Institution: | (1) Department of Agriculture and Food Western Australia, 3 Baron-Hay Court, South Perth, 6151, Australia |
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Abstract: | A key challenge in marker-assisted selection (MAS) for molecular plant breeding is to develop markers linked to genes of interest
which are applicable to multiple breeding populations. In this study representative F2 plants from a cross Mandalup (resistant to anthracnose disease) × Quilinock (susceptible) of Lupinus angustifolius were used in DNA fingerprinting by Microsatellite-anchored Fragment Length Polymorphism (MFLP). Nine candidate MFLP markers
linked to anthracnose resistance were identified, then ‘validated’ on 17 commercial cultivars. The number of “false positives”
(showing resistant-allele band but lack of the R gene) for each of the nine candidate MFLP markers on the 17 cultivars ranged
from 1 to 9. The candidate marker with least number of false positive was selected, sequenced, and was converted into a co-dominant,
sequence-specific, simple PCR based marker suitable for routine implementation. Testing on 180 F2 plants confirmed that the converted marker was linked to the R gene at 5.1 centiMorgan. The banding pattern of the converted
marker was consistent with the disease phenotype on 23 out of the 24 cultivars. This marker, designated “AnManM1”, is now
being used for MAS in the Australian lupin breeding program. We conclude that generation of multiple candidate markers, followed
by a validation step to select the best marker before conversion to an implementable form is an efficient strategy to ensure
wide applicability for MAS. |
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Keywords: | Marker assisted selection (MAS) Molecular marker MFLP Lupinus angustifolius |
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