Efficient generation of targeted point mutations in the Brassica oleracea var. botrytis genome via a modified CRISPR/Cas9 system
文献类型： 外文期刊
作者： Wang, Guixiang 1 ; Zong, Mei 1 ; Liu, Di 1 ; Wu, Yage 1 ; Tian, Shouwei 1 ; Han, Shuo 1 ; Guo, Ning 1 ; Duan, Mengmeng 1 ; Miao, Liming 1 ; Liu, Fan 1 ;
作者机构： 1.Beijing Acad Agr & Forestry Sci, Beijing Vegetable Res Ctr, Key Lab Biol & Genet Improvement Hort Crops North, Beijing Key Lab Vegetable Germplasm Improvement,Mi, Beijing 100097, Peoples R China
关键词： Cauliflower; Targeted point mutations; Base-editing; CRISPR; Cas9; ALS; CENH3
期刊名称：HORTICULTURAL PLANT JOURNAL （ 影响因子：4.24； ）
ISSN： 2095-9885
年卷期： 2022 年 8 卷 4 期
页码：
收录情况： SCI
摘要： A B S T R A C T 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.
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