Abstract
SummaryMembers 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 T2 generation in self‐pollinating species. Analysis of dry weights and flowering times for four independent T3 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.
Highlights
Plant viruses are ubiquitous in natural environments and can severely limit plant growth and fertility
The Potyvirus genus contains a greater number of virus species than any other plant virus genus (Gibbs and Ohshima, 2010), and certain species within this genus [notably its type member, Potato virus Y (PVY)] are damaging to economically important crops (Karasev and Gray, 2013)
As loss-of-function mutations in components of the eIF4F translation complex have repeatedly been associated with stable resistance to several potyviruses, we aimed to generate virusresistant plants by novel mutation at the eukaryotic translation initiation factor (eIF)(iso)4E locus in Arabidopsis thaliana using CRISPR/Cas9 technology
Summary
Plant viruses are ubiquitous in natural environments and can severely limit plant growth and fertility. Viruses are a significant economic burden to both well-developed and underdeveloped agriculture because of absolute yield losses in the field and decreased marketability of harvested crops. The Potyvirus genus contains a greater number of virus species than any other plant virus genus (Gibbs and Ohshima, 2010), and certain species within this genus [notably its type member, Potato virus Y (PVY)] are damaging to economically important crops (Karasev and Gray, 2013). Potyviruses exist as flexuous, rod-shaped virions comprising a positive-sense, single-stranded RNA (1ssRNA) genome coated by a virally encoded coat protein (Jagadish et al, 1991). On entry into plant cells, the 1ssRNA genome is uncoated and translated into a single polypeptide, which subsequently generates the range of potyviral proteins by autocatalysis (Carrington et al, 1989). Translation of the potyviral RNA is largely dependent on host translation factors
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