Journal
SCIENTIA HORTICULTURAE
Volume 319, Issue -, Pages -Publisher
ELSEVIER
DOI: 10.1016/j.scienta.2023.112136
Keywords
Broccoli; CRISPR; Cas9; BoGL5; Functional verification
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CRISPR/Cas9 system is utilized for precise gene editing in broccoli, a globally important vegetable, by introducing multiple single-guide RNA-expressing cassettes. The study demonstrates that CRISPR/Cas9 can be a promising tool for gene functional verification and trait improvement in broccoli.
CRISPR/Cas9 has been widely used for precise and efficient genome editing in a number of organisms, notably in major crops, for gene function characterization and trait improvement. However, the application of this genome editing tool is very limited in Brassica oleracea, especially in broccoli (Brassica oleracea var. italica). Here, we report that the genome of broccoli, a worldwide important Brassicaceae vegetable, can also be precisely edited by a CRISPR/Cas9 system with multiple single-guide RNA-expressing cassettes. The gene BoGL5, previously identified as a candidate for the glossy green mutant, was chosen as the target gene. We generated 14 T0 transgenic plants, among which 14.3% and 50% produced mutations at the expected position in targeted regions 1 and 2, respectively. Two of them with a 100% mutation rate showed a glossy green phenotype and dramatically reduced cuticular wax load. These results indicate that CRISPR/Cas9 can be used as a promising technique for gene functional verification and trait improvement in broccoli.
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