Engineering of CRISPR/Cas9‐mediated potyvirus resistance in transgene‐free Arabidopsis plants

Abstract

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.

Cited by papers we hold 13

• CRISPR-Cas-Led Revolution in Diagnosis and Management of Emerging Plant Viruses: New Avenues Toward Food and Nutritional Security · 2021
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• Detection of Plant Viruses and Disease Management: Relevance of Genetic Diversity and Evolution · 2020
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• Efficient and modular CRISPR‐Cas9 vector system for Physcomitrella patens · 2019
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• Mimicking natural polymorphism in eIF 4E by CRISPR ‐Cas9 base editing is associated with resistance to potyviruses · 2019
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• The Enhancement of Plant Disease Resistance Using CRISPR/Cas9 Technology · 2018
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• CRISPR for Crop Improvement: An Update Review · 2018
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• Novel alleles of rice eIF4G generated by CRISPR/Cas9‐targeted mutagenesis confer resistance to Rice tungro spherical virus · 2018
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• Simultaneous CRISPR /Cas9‐mediated editing of cassava eIF 4E isoforms nCBP ‐1 and nCBP ‐2 reduces cassava brown streak disease symptom severity and incidence · 2018
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• CRISPR/Cas9‐mediated efficient targeted mutagenesis in grape in the first generation · 2017
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• CRISPR/Cas9‐mediated targeted mutagenesis of GmFT2a delays flowering time in soya bean · 2017
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• Knockout of OsNramp5 using the CRISPR/Cas9 system produces low Cd-accumulating indica rice without compromising yield · 2017
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• Plant genome editing with TALEN and CRISPR · 2017
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• Gene Editing and Crop Improvement Using CRISPR-Cas9 System · 2017
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