Efficient CRISPR/Cas-Mediated Targeted Mutagenesis in Spring and Winter Wheat Varieties

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.

Advances in understanding the soil-borne viruses of wheat: from the laboratory bench to strategies for disease control in the field

“…In cucumbers, knocking out the eIF4E gene using the CRISPR/Cas9 system resulted in broad-spectrum resistance to members of the Potyviridae family, including cucumber vein yellowing virus, papaya ringspot virus, and zucchini yellow mosaic virus (Chandrasekaran et al 2016). In wheat, TaeIF(iso)4e-mutant lines were obtained using CRISPR-Cas9 genome-editing technology, which revealed that the corresponding mutations can potentially confer WSSMV and WYMV resistance (Hahn et al 2021). Recently, the wheat orthologs of the barley susceptibility factor disulfide isomerase like 5-1 (HvPDIL5-1) were edited in the wheat genome, conferring WYMV resistance in wheat (Kan et al 2022).…”

Advances in Editing Silkworms (Bombyx mori) Genome by Using the CRISPR-Cas System

“…CRISPR-Cas is one of the key methods employed by many molecular biology scientific laboratories. Since its first description [14], genome editing focused research was implemented by countless research groups [15][16][17].…”