Enriching Vitamin C in Tomato Fruits by CRISPR/Cas9-Mediated SlCSN5B Gene Editing
10.13865/j.cnki.cjbmb.2025.01.1412
- VernacularTitle:基于CRISPR/Cas9技术创制高维生素C番茄材料
- Author:
Xuan-Sen AN
1
;
Yan-Wei WANG
1
;
Wen-Chao TIAN
1
;
Ya-Juan REN
1
;
Peng-Fei AI
1
Author Information
1. 河北科技大学食品与生物学院,石家庄 050018
- Publication Type:Journal Article
- Keywords:
clustered regularly interspaced short palindromic repeats/CRISPR-associated(Cas)sys-tems 9(CRISPR/Cas9);
tomato;
gene encoding subunit 5B of constitutively photomorphogenic 9 signa-losome in tomato(SlCSN5B);
vitamin C(Vit C)
- From:
Chinese Journal of Biochemistry and Molecular Biology
2025;41(3):460-469
- CountryChina
- Language:Chinese
-
Abstract:
Subunit 5B of constitutively photomorphogenic 9 signalosome(CSN5B)is an inhibitory factor for the biosynthesis of vitamin C in the L-galactose synthesis pathway in plants.To create mutants with richer vitamin C in tomato fruits,a dual-target vector of pKSE402-SlCSN5Bwas constructed and trans-formed into the breeding parent of 1912.Based on the molecular biological assay and DNA sequencing,17 transgenic positive lines were determined,and 6 lines of them were genetically mutated at the target site of SlCSN5B,with an editing efficiency of 35.3％.Among the mutants,2 lines were homozygous mu-tants,csn5b-6 with 180 bp deletion and csn5b-8 with 3 bp deletion.The biological traits of two types of deficiency homozygotes without exogenous T-DNA insertion derived from the T1 generation were observed,and there were no significant differences in phenotypes of the plant and fruit.Through physiological tes-ting of red-ripe fruits from T,generation lines of csn5b-6,the GMPase activity and the vitamin C content significantly increased by 43％and 37.8％,respectively,the content of hydrogen peroxide significantly decreased by 25.9％,and the content of soluble solids did not obviously change,compared with the wild type.There were no significant differences in plant traits and physiological characteristics between T1 gen-eration lines of csn5b-8 and the wild type.Based on gene expression analysis and protein structure predic-tion,the results showed that the gene of SlCSN5B normally transcribed,and the loss of large peptide seg-ments of CSN5B encoded by SlCSN5B caused changes in its structure to affect functioning,which led to an increase of vitamin C content in the line of csn5b-6-1 1.The findings suggest that the vitamin C content of tomato fruit can be improved by CRISPR/Cas9-mediated SlCSN5B gene editing,which provides the valuable resources for high-quality breeding in tomato.
