GE-WRKY70 canola lines were generated by CRISPR-Cas editing of the transcription factor gene wrky70 in the allotetraploid Brassica napus line J9712.
WRKY transcription factors are widely involved in defense to diverse plant stress conditions. wrky70 seems to be involved in the response to pathogens in B. napus.
Aim of the mutation of the wrky70 gene was the improvement of canola resistance to Sclerotinia sclerotiorum.
A CRISPR/Cas9 vector with multiple sgRNA expression cassettes targeting the first exon of four wrky70 homoeologs, and the selectable npt gene was designed to generate WRKY70 mutants. Agrobacterium-mediated genetic transformation was used to generate transgenic plants.
The mutations of targeted sites were investigated by amplifying and sequencing in the T0 and T1 generations. Analysis of the mutation pattern revealed insertions of mostly 1 bp and deletions.
T1 plants with three mutated copies of wrky70 exhibited enhanced resistance to S. sclerotiorum, when compared to non-transgenic plants.
Source:
Sun et al. (2018) CRISPR/Cas9-Mediated Multiplex Genome Editing of the BnWRKY11 and BnWRKY70 Genes in Brassica napus L., Int. J. Mol. Sci., 19, 2716; doi:10.3390/ijms19092716
