Journal
MOLECULAR PLANT PATHOLOGY
Volume 17, Issue 8, Pages 1276-1288Publisher
WILEY
DOI: 10.1111/mpp.12417
Keywords
CRISPR; Cas9; eIF(iso)4E; potyvirus; transgene-free; TuMV; virus resistance
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Funding
- Biotechnology and Biological Sciences Research Council (BBSRC) EastBio Doctoral Training Partnership (DTP) [BB/J01446X/1]
- James Hutton Institute studentship
- Biotechnology and Biological Sciences Research Council [1311408] Funding Source: researchfish
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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.
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