Rapid cloning of disease-resistance genes in plants using mutagenesis and sequence capture.
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.
The Sainsbury Laboratory, Norwich, UK.|John Innes Centre, Norwich, UK.
Burkhard
B
Steuernagel
Commonwealth Scientific and Industrial Research Organization (CSIRO), Agriculture Flagship, Canberra, NSW, Australia.
Sambasivam K
SK
Periyannan
The Sainsbury Laboratory, Norwich, UK.
Inmaculada
I
Hernández-Pinzón
The Sainsbury Laboratory, Norwich, UK.
Kamil
K
Witek
USDA-ARS Cereal Disease Laboratory and Department of Plant Pathology, University of Minnesota, St. Paul, Minnesota, USA.
Matthew N
MN
Rouse
John Innes Centre, Norwich, UK.
Guotai
G
Yu
John Innes Centre, Norwich, UK.|Department of Agriculture Technology, Universiti Putra Malaysia, Serdang, Malaysia.
Asyraf
A
Hatta
Commonwealth Scientific and Industrial Research Organization (CSIRO), Agriculture Flagship, Canberra, NSW, Australia.
Mick
M
Ayliffe
University of Sydney, Plant Breeding Institute, Cobbitty, NSW, Australia.
Harbans
H
Bariana
The Sainsbury Laboratory, Norwich, UK.
Jonathan D G
JD
Jones
Commonwealth Scientific and Industrial Research Organization (CSIRO), Agriculture Flagship, Canberra, NSW, Australia.
Evans S
ES
Lagudah
The Sainsbury Laboratory, Norwich, UK.|John Innes Centre, Norwich, UK.
Brande B H
BB
Wulff
D003001:Cloning, Molecular / Q000379:methods*; D060467:Disease Resistance / Q000235:genetics*; D017343:Genes, Plant / Q000235:genetics*; D024861:Genetic Enhancement / Q000379:methods; D016297:Mutagenesis, Site-Directed / Q000379:methods*; D010935:Plant Diseases / Q000235:genetics* / Q000517:prevention & control; D010944:Plants / Q000235:genetics*; D030821:Plants, Genetically Modified / Q000235:genetics; D017422:Sequence Analysis, DNA / Q000379:methods