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Nat Biotechnol. 2016 Jun;34(6):652-5. doi: 10.1038/nbt.3543. Epub 2016 Apr 25.

Rapid cloning of disease-resistance genes in plants using mutagenesis and sequence capture.

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The Sainsbury Laboratory, Norwich, UK.
John Innes Centre, Norwich, UK.
Commonwealth Scientific and Industrial Research Organization (CSIRO), Agriculture Flagship, Canberra, NSW, Australia.
USDA-ARS Cereal Disease Laboratory and Department of Plant Pathology, University of Minnesota, St. Paul, Minnesota, USA.
Department of Agriculture Technology, Universiti Putra Malaysia, Serdang, Malaysia.
University of Sydney, Plant Breeding Institute, Cobbitty, NSW, Australia.


Wild relatives of domesticated crop species harbor multiple, diverse, disease resistance (R) genes that could be used to engineer sustainable disease control. However, breeding R genes into crop lines often requires long breeding timelines of 5-15 years to break linkage between R genes and deleterious alleles (linkage drag). Further, when R genes are bred one at a time into crop lines, the protection that they confer is often overcome within a few seasons by pathogen evolution. If several cloned R genes were available, it would be possible to pyramid R genes in a crop, which might provide more durable resistance. We describe a three-step method (MutRenSeq)-that combines chemical mutagenesis with exome capture and sequencing for rapid R gene cloning. We applied MutRenSeq to clone stem rust resistance genes Sr22 and Sr45 from hexaploid bread wheat. MutRenSeq can be applied to other commercially relevant crops and their relatives, including, for example, pea, bean, barley, oat, rye, rice and maize.

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