Open AccessEngineering of CRISPR/Cas9‐mediated potyvirus resistance in transgene‐free Arabidopsis plants
Author(s) -
Pyott Douglas E.,
Sheehan Emma,
Molnar Attila
Publication year - 2016
Publication title -
molecular plant pathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.945
H-Index - 103
eISSN - 1364-3703
pISSN - 1464-6722
DOI - 10.1111/mpp.12417
Subject(s) - biology , crispr , potyvirus , genetics , transgene , potato virus y , cas9 , genetically modified crops , locus (genetics) , gene , arabidopsis , allele , potyviridae , turnip mosaic virus , plant disease resistance , mutant , virology , plant virus , virus
Summary Members of the eukaryotic translation initiation factor (eIF) gene family, including eIF4E and its paralogue eIF(iso)4E , have previously been identified as recessive resistance alleles against various potyviruses in a range of different hosts. However, the identification and introgression of these alleles into important crop species is often limited. In this study, we utilise CRISPR/Cas9 technology to introduce sequence‐specific deleterious point mutations at the eIF(iso)4E locus in Arabidopsis thaliana to successfully engineer complete resistance to Turnip mosaic virus (TuMV), a major pathogen in field‐grown vegetable crops. By segregating the induced mutation from the CRISPR/Cas9 transgene, we outline a framework for the production of heritable, homozygous mutations in the transgene‐free T 2 generation in self‐pollinating species. Analysis of dry weights and flowering times for four independent T 3 lines revealed no differences from wild‐type plants under standard growth conditions, suggesting that homozygous mutations in eIF(iso)4E do not affect plant vigour. Thus, the established CRISPR/Cas9 technology provides a new approach for the generation of Potyvirus resistance alleles in important crops without the use of persistent transgenes.