Efficient BoPDS Gene Editing in Cabbage by the CRISPR/Cas9 System

Abstract

Genome editing offers great advantages in identifying gene function and generating agronomically important mutations in crops. Here, we report that the genome of cabbage (Brassica oleracea var. capitata), an important cruciferous vegetable, can also be precisely edited by a CRISPR/Cas9 system stacked with multiple single-guide RNA-expressing cassettes. When the phytoene desaturase BoPDS gene was used as the target gene, an albino-phenotype transgenic shoot in T0 Basta-resistant lines was observed, and 37.5% of the transgenic cabbage shoots carried BoPDS gene mutations as a result of nucleotide deletions at the expected position. Moreover, mutations were detected in sites with the same target sequence in gene Bol016089 which is paralogous to the BoPDS gene. Our results show that the CRISPR/ Cas9 system is a powerful tool for cabbage variety improvement by genome editing. Keywords: Cabbage, CRISPR/Cas9, BoPDS, Genome editing, Mutation