Simultaneous editing of three homoeoalleles in hexaploid bread wheat confers heritable resistance to powdery mildew

Sequence-specific nucleases have been applied to engineer targeted modifications in polyploid genomes, but simultaneous modification of multiple homoeoalleles has not been reported. Here we use transcription activator-like effector nuclease (TALEN) and clustered, regularly interspaced, short palindr...

Full description

Saved in:
Bibliographic Details
Published in:Nature biotechnology 2014-09, Vol.32 (9), p.947-951
Main Authors: Wang, Yanpeng, Cheng, Xi, Shan, Qiwei, Zhang, Yi, Liu, Jinxing, Gao, Caixia, Qiu, Jin-Long
Format: Article
Language:English
Subjects:
Alleles
Allelomorphism
Ascomycota - genetics
Ascomycota - pathogenicity
Base Sequence
Bread
Diseases and pests
Genetic aspects
Genomes
Health aspects
Mildew
Molecular Sequence Data
Mutation
Plant immunology
Proteins
RNA Editing
Sequence Homology, Nucleic Acid
Transgenic plants
Triticum - genetics
Triticum - microbiology
Triticum aestivum
Wheat
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Sequence-specific nucleases have been applied to engineer targeted modifications in polyploid genomes, but simultaneous modification of multiple homoeoalleles has not been reported. Here we use transcription activator-like effector nuclease (TALEN) and clustered, regularly interspaced, short palindromic repeats (CRISPR)-Cas9 (refs. 4,5) technologies in hexaploid bread wheat to introduce targeted mutations in the three homoeoalleles that encode MILDEW-RESISTANCE LOCUS (MLO) proteins. Genetic redundancy has prevented evaluation of whether mutation of all three MLO alleles in bread wheat might confer resistance to powdery mildew, a trait not found in natural populations. We show that TALEN-induced mutation of all three TaMLO homoeologs in the same plant confers heritable broad-spectrum resistance to powdery mildew. We further use CRISPR-Cas9 technology to generate transgenic wheat plants that carry mutations in the TaMLO-A1 allele. We also demonstrate the feasibility of engineering targeted DNA insertion in bread wheat through nonhomologous end joining of the double-strand breaks caused by TALENs. Our findings provide a methodological framework to improve polyploid crops.
ISSN:1087-0156
1546-1696
DOI:10.1038/nbt.2969