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

Area of Science:

• Plant genetics and molecular biology
• Crop improvement and biotechnology

Background:

• Wheat (Triticum aestivum) is a vital global crop with significant potential for trait improvement using genome editing.
• CRISPR/Cas technology is a powerful tool for gene function studies and crop enhancement in plants.

Purpose of the Study:

• To utilize CRISPR/Cas technology for genome editing in wheat, targeting specific genes.
• To generate mutant lines for studying the roles of BAK1 in immunity and Ta-eIF4E/Ta-eIF(iso)4E in viral susceptibility.

Main Methods:

• CRISPR/Cas-induced mutations (indels) were generated in three target genes: TaBAK1-2, Ta-eIF4E, and Ta-eIF(iso)4E.
• Experiments were conducted on both spring (Cadenza) and winter (Cezanne, Goncourt, Prevert) wheat cultivars.
• Successful generation of primary transgenic lines with CRISPR/Cas-induced indels was achieved for all targeted genes.

Main Results:

• Primary transgenic lines with CRISPR/Cas-induced indels were successfully generated for TaBAK1-2, Ta-eIF4E, and Ta-eIF(iso)4E in wheat.
• The study demonstrated effective genome editing in both spring and winter wheat cultivars.
• The methodology provides a foundation for future research in wheat genetic improvement.

Conclusions:

• Homozygous tabak1-2 mutant lines will aid research into BAK1's role in wheat immune responses.
• The ta-eif4e and ta-eif(iso)4e mutants hold potential for developing resistance against wheat spindle streak mosaic virus (WSSMV) and wheat yellow mosaic virus (WYMV).
• The presented transformation and genome editing methods are valuable for the research community, particularly for winter wheat.