Next-generation sequencing of amplicons is a rapid and reliable method for the detection of polymorphisms relevant for barley breeding
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Modern plant breeding is based on the detection of traits at the molecular level so that the selection process can be accelerated. Methods that allow the rapid and high-throughput analysis of such traits are therefore useful for plant breeders and farmers. We have shown that next-generation sequencing, applied to short PCR products spanning or flanking polymorphisms of interest in barley, can provide robust genotyping data that allow the rapid determination of genotype and zygosity. This method can be used to genotype large panels of plants because up to 80 million individual reads can be produced in one sequencing run, and samples from different lines and/or traits can be pooled. These findings are significant because plant breeders may need to screen large populations for multiple traits in parallel. Our next-generation sequencing method therefore provides a simple and inexpensive approach for the rapid and accurate genotyping of natural polymorphisms in barley, which can also be applied in many other economically relevant crop species.
KeywordsCandidate gene Crops High-throughput genotyping Polymorphism QTL SNP
We acknowledge Harold E. Bockelman from the United States Department of Agriculture (USDA) for providing the barley seeds. We acknowledge Cornelia Marthe and Dr. Jochen Kumlehn (IPK Gatersleben, Group Plant Reproductive Biology) for sharing their expertise on the crossbreeding of barley. We acknowledge Dr. Richard M Twyman for editing the manuscript. This work is supported by the BMBF (0316075B).
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