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ARS Home » Midwest Area » St. Paul, Minnesota » Cereal Disease Lab » Research » Publications at this Location » Publication #322809
Title: Rapid cloning of disease-resistance genes in plants using mutagenesis and sequence capture

Author
item STEUERNAGEL, BURKHARD - John Innes Center
item PERIYANNAN, SAMBASIVAM - Commonwealth Scientific And Industrial Research Organisation (CSIRO)
item HERNANDEZ-PINZON, INMACULADA - Sainsbury Laboratory
item WITEK, KAMIL - Sainsbury Laboratory
item Rouse, Matthew - Matt
item YU, GUOTAI - John Innes Center
item HATTA, ASYRAF - John Innes Center
item AYLIFFE, MICK - Commonwealth Scientific And Industrial Research Organisation (CSIRO)
item BARIANA, HARBANS - University Of Sydney
item JONES, JONATHAN - Sainsbury Laboratory
item LAGUDAH, EVANS - Commonwealth Scientific And Industrial Research Organisation (CSIRO)
item WULFF, BRANDE - John Innes Center

Submitted to: Nature Biotechnology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 3/16/2016
Publication Date: 4/25/2016
Citation: Steuernagel, B., Periyannan, S., Hernandez-Pinzon, I., Witek, K., Rouse, M.N., Yu, G., Hatta, A., Ayliffe, M., Bariana, H., Jones, J., Lagudah, E., Wulff, B. 2016. Rapid cloning of disease-resistance genes in plants using mutagenesis and sequence capture. Nature Biotechnology. 34:652-655. doi:10.1038/nbt.3543.

Interpretive Summary: Emerging virulent races of the wheat stem rust pathogen threaten food security. Genetic solutions to protect wheat from stem rust are needed. Wild crop relatives carry disease resistance (R) genes. However, recruiting R genes for crop improvement typically involves long breeding timelines and R genes are often overcome by pathogen adaptation within a few seasons when deployed one at a time. With cloned R genes, we could speed up disease resistance breeding and simultaneously introduce multiple R genes, which should delay resistance-breaking pathogen race evolution and provide more durable resistance. We describe a three-step method (MutRenSeq) for rapid R gene cloning based on (i) chemical mutagenesis and screening for susceptible mutants, (ii) exome capture and sequencing of R gene family members (iii) sequence comparison of wild type and mutants. We use MutRenSeq to clone the important stem rust resistance genes Sr22 and Sr45 from hexaploid bread wheat. Cloning multiple stem rust resistance genes will enable breeding of United States wheat cultivars with durable resistance to stem rust.

Technical Abstract: Genetic solutions to protect crops against pests and pathogens are preferable to agrichemicals 1. Wild crop relatives carry immense diversity of disease resistance (R) genes that could enable more sustainable disease control. However, recruiting R genes for crop improvement typically involves long breeding timelines to break linkage to deleterious alleles of other genes (“linkage drag”), and R genes are often overcome within a few seasons when deployed one at a time2. With cloned R genes, we could avoid linkage drag and simultaneously introduce multiple R genes3, which should delay resistance-breaking pathogen race evolution and provide more durable resistance2. We describe a three-step method (MutRenSeq) for rapid R gene cloning based on (i) chemical mutagenesis and screening for susceptible mutants, (ii) exome capture and sequencing of R gene family members (iii) sequence comparison of wild type and mutants. We used MutRenSeq to clone the important stem rust resistance genes Sr22 and Sr45 from hexaploid bread wheat.