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Development of sequence-specific PCR markers associated with a polygenic controlled trait for marker-assisted selection using a modified selective genotyping strategy: a case study on anthracnose disease resistance in white lupin (Lupinus albus L.)

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Abstract

Selection for anthracnose disease resistance is one of the top priorities in white lupin (Lupinus albus) breeding programs. A cross was made between a landrace P27174 (resistant to anthracnose) and a cultivar Kiev Mutant (susceptible). The progeny was advanced to F8 recombinant inbred lines (RILs). Disease tests on the RIL population from field trials over 2 years indicated that the disease resistance in P27174 was polygenic controlled. A modified selective genotyping strategy was applied in the development of molecular markers linked to quantitative loci conferring anthracnose diseases resistance. Eight individual plants representing high level of anthracnose resistance (HR), eight plants representing susceptibility (S), together with eight lines representing medium level of anthracnose resistance (MR), were subjected to DNA fingerprinting by Microsatellite-anchored Fragment Length Polymorphisms (MFLP). Six MFLP polymorphisms, which had the banding pattern matching the HR plants and the S plants, were identified as candidate markers linked to quantitative loci conferring anthracnose resistance. The six candidate MFLP markers were delineated into three groups based on their banding variation on the eight MR plants. One candidate MFLP marker each from the three groups was selected, cloned, sequenced, and converted into co-dominant, sequence-specific PCR markers. These three markers, designated as WANR1, WANR2 and WANR3, were tested on a segregating population containing 189 F8 RILs. The disease phenotyping data and the marker genotyping data on the F8 RILs were merged and analysed by the JMP software using the ‘fit-model’ function, which revealed that 71% of the phenotypic variation was controlled by genetic factors, while the other 29% of the phenotypic variation was due to environmental factors and environment × genotype interactions. On individual marker basis, marker WANR1 conditioned 39% of phenotypic variations of anthracnose resistance, followed by marker WANR2 with 8%, and WANR3 with 12%. Further analysis showed that WANR2 and WANR3 were on the same linkage group with a genetic distance of 15.3 cM. The combination of the two markers WANR1 and WANR3 explained 51% out from the 71% of the genetic controlled variations for disease resistance, indicating that the two QTLs working additively for anthracnose disease resistance. A simulation of marker-assisted selection on the F8 RIL population using the two markers WANR1 and WANR3 identified 42 out of the 189 RILs being homozygous for resistance-allele bands for both markers, and 41 of them showed disease severity below 3.0 on the 1 (highly resistant) to 5 (susceptible) scale. The two markers WANR1 and WANR3 have now been implemented for marker-assisted selection for anthracnose resistance in the L. albus breeding program in Australia.

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Acknowledgments

This research was supported by the Grains Research and Development Corporation (GRDC), Australia. We thank Dr. Simon Ellwood for the help in making comparison of our marker results with the results reported in Phan et al. (2007).

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Authors and Affiliations

  1. Department of Agriculture and Food Western Australia, 3 Baron-Hay Court, South Perth, WA, 6151, Australia

    Huaan Yang, Daniel Renshaw, Chengdao Li, Kedar Adhikari, Geoff Thomas, Bevan Buirchell & Mark Sweetingham

  2. State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, 100193, Beijing, People’s Republic of China

    Ruiming Lin

  3. School of Plant Biology, Faculty of Natural and Agricultural Sciences, The University of Western Australia, 35 Stirling Highway, Crawley, WA, 6009, Australia

    Guijun Yan

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  1. Huaan Yang
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  2. Ruiming Lin
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  3. Daniel Renshaw
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  4. Chengdao Li
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  5. Kedar Adhikari
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  6. Geoff Thomas
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  7. Bevan Buirchell
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  8. Mark Sweetingham
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  9. Guijun Yan
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Correspondence to Huaan Yang.

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Yang, H., Lin, R., Renshaw, D. et al. Development of sequence-specific PCR markers associated with a polygenic controlled trait for marker-assisted selection using a modified selective genotyping strategy: a case study on anthracnose disease resistance in white lupin (Lupinus albus L.). Mol Breeding 25, 239–249 (2010). https://doi.org/10.1007/s11032-009-9325-4

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