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Efficient Generation of Targeted Point Mutations in the Brassica oleracea var. botrytis Genome via a Modified CRISPR/Cas9 System
Horticultural Plant Journal  (IF3.032),  Pub Date : 2022-01-21, DOI: 10.1016/j.hpj.2022.01.005
Guixiang Wang, Mei Zong, Di Liu, Yage Wu, Shouwei Tian, Shuo Han, Ning Guo, Mengmeng Duan, Liming Miao, Fan Liu

In this study, we used the modified CRISPR/Cas9 system to produce targeted point mutations in cauliflower. Acetolactate synthase (ALS) and Centromere-specific histone H3 variant (CENH3) genes were selected as the base-editing targets and hypocotyls of cauliflower were used as explants. For ALS gene, a C-to-T conversion in the Pro182 codon (CCT) can alter the encoded amino acid, likely resulting in herbicide resistance, and a C-to-T mutation in the Leu133 codon (CTT) in the CENH3 gene may produce a haploid inducer. Results indicated that the transformation efficiency was 1.8%–4.5% and the mutation efficiencies for the ALS and CENH3 genes were approximately 22% and 87%, respectively. The ALS mutant cauliflower showed strong herbicide resistance, with possible immediate implications for broadleaf weed control in cauliflower fields.