Development and validation of a breeder-friendly KASPar marker for wheat leaf rust resistance locus Lr21
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Development and utilization of genetic markers play a pivotal role in marker-assisted breeding of wheat cultivars with pyramids of disease resistance genes. The objective of this study was to develop a closed-tube, gel-free assay for high-throughput genotyping of leaf rust resistance locus Lr21. Polymorphism identified from re-sequencing of a 2.4-kb fragment covering the functional region of the Lr21 gene from the second to the fourth indels was targeted for assay development. The generated sequence data revealed the 88- or 105-bp indel in the first intron of the Lr21 gene in the selected resistant cultivars compared to susceptible US spring and winter wheat cultivars. Allele-specific primers for a KASPar assay were designed around the junction of the indel at position 1,346 bp. The marker was tested on a panel of 384 US wheat lines and found to be effective in differentiating resistant and susceptible genotypes.
KeywordsLeaf rust KASPar assay SNP genotyping Puccinia triticina Eriks.
The contribution of Mr. Jared Smith and Mrs. Sharon Williamson (USDA-ARS Eastern Regional Small Grains Genotyping, Laboratory) in designing KASPar assays and assay setup is acknowledged with thanks. This work was supported in part by funds provided by the USDA T-CAP AFRI grant number 2011-68002-30029.
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