Abstract

CRISPR/Cas technology has recently become the molecular tool of choice for gene function studies in plants as well as crop improvement. Wheat is a globally important staple crop with a well annotated genome and there is plenty of scope for improving its agriculturally important traits using genome editing technologies, such as CRISPR/Cas. As part of this study we targeted three different genes in hexaploid wheat Triticum aestivum: TaBAK1-2 in the spring cultivar Cadenza as well as Ta-eIF4E and Ta-eIF(iso)4E in winter cultivars Cezanne, Goncourt and Prevert. Primary transgenic lines carrying CRISPR/Cas-induced indels were successfully generated for all targeted genes. While BAK1 is an important regulator of plant immunity and development, Ta-eIF4E and Ta-eIF(iso)4E act as susceptibility (S) factors required for plant viruses from the Potyviridae family to complete their life cycle. We anticipate the resultant homozygous tabak1-2 mutant lines will facilitate studies on the involvement of BAK1 in immune responses in wheat, while ta-eif4e and ta-eif(iso)4e mutant lines have the potential to become a source of resistance to wheat spindle streak mosaic virus (WSSMV) and wheat yellow mosaic virus (WYMV), both of which are important pathogens of wheat. As winter wheat varieties are generally less amenable to genetic transformation, the successful experimental methodology for transformation and genome editing in winter wheat presented in this study will be of interest to the research community working with this crop.

Highlights

  • Common wheat (Triticum aestivum) is one of the most important staple food crops in the world

  • During this study we generated tabak1-2 lines carrying different combinations of mutant tabak1-2a, tabak1-2b and tabak1-2d alleles, including multiple homozygous lines (Figure S7), in the spring wheat cultivar Cadenza, which is relatively easy to transform and whose genome has been sequenced as part of the 10+ Wheat Genomes Project

  • BRI1associated receptor kinase 1 (BAK1) acts as a coreceptor for a number of pattern recognition receptors (PRRs) involved in pattern-triggered immunity (PTI) responses in plants [5]

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Summary

Introduction

Common wheat (Triticum aestivum) is one of the most important staple food crops in the world. The challenges that global agriculture currently faces, such as growth of the world’s population and climate change, dictate demand for technologies with a potential to accelerate crop breeding [1]. Genome editing emerged as a powerful new breeding technique (NBT) [2] that enables targeted changes in crop genomes. CRISPR/Cas is by far the most common plant genome editing technology nowadays due to its precision, versatility and ease of use [3]. It is an excellent tool for gene function studies as well as improvement of agriculturally important crop traits. The CRISPR/Cas technology has been successfully used for both above-mentioned applications

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