Efficient Editing of SoCSLD2 by CRISPR/Cas9 Affects Morphogenesis of Root Hair in Spinach
文献类型： 外文期刊
作者： Cao, Yingping 1 ; Xu, Yue 2 ; Zhang, Yue 1 ; Zhang, Heng 1 ; Qin, Zhi 1 ; Bai, Chen 1 ; Zhang, Hailing 5 ; Ma, Dongmei 6 ; Wang, Quanhua 1 ; Fu, Chunxiang 2 ; Sun, Meihong 1 ; Dai, Shaojun 1 ;
作者机构： 1.Shanghai Normal Univ, Coll Life Sci, Dev Ctr Plant Germplasm Resources, Shanghai 200234, Peoples R China
2.Chinese Acad Sci, Shandong Prov Key Lab Energy Genet, Shandong Technol Innovat Ctr Synthet Biol, Qingdao 266000, Peoples R China
3.Chinese Acad Sci, Qingdao Inst Bioenergy & Bioproc Technol, CAS Key Lab Biofuels, Qingdao 266000, Peoples R China
4.Northeast Forestry Univ, Coll Life Sci, Minist Educ, Key Lab Saline Alkali Vegetat Ecol Restorat, Harbin 150040, Peoples R China
5.Heilongjiang Acad Agr Sci, Grass & Sci Inst, Harbin 150086, Peoples R China
6.Ningxia Univ, Sch Ecol & Environm, Breeding Base State Key Lab Land Degradat & Ecol, Key Lab Restorat & Reconstruct Degraded Ecosyst N, Yinchuan 750021, Peoples R China
关键词： CRISPR; Cas9; cellulose synthase-like D (CSLD) gene; spinach; hairy root; root hair; transcriptomic analysis
期刊名称：HORTICULTURAE （ 影响因子：3.0； 五年影响因子：3.2 ）
ISSN：
年卷期： 2022 年 8 卷 8 期
页码：
收录情况： SCI
摘要： CRISPR/Cas9 is a valuable tool and has been extensively employed to perform gene editing in plants. However, CRISPR/Cas9 has not been successfully used in spinach, an important leafy vegetable crop. Here, we established a CRISPR/Cas9-based gene-editing system for spinach hairy roots and edited two cellulose synthase-like D (CSLD) genes (SoCSLD2 and SoCSLD3) that were involved in root-hair formation of spinach hairy roots. Four mutation types (i.e., replacement, insertion, deletion, and combined mutations) were observed, among which the replacement accounted for the vast majority (about 64.1%). Mutation rate differed largely among different targets. Seven homozygous/bi-allelic and eight heterozygous/chimeric mutants of SoCSLD2 were obtained from 15 independent transgenic hairy root lines. All of the seven homozygous/bi-allelic mutant lines displayed bulking and short root hairs, which resembled the characteristics of Arabidopsis atcsld2 mutants. The transcriptomic analysis further revealed that multiple gene expressions for cell-wall modulation and membrane trafficking were disturbed, which might result in the inhibition of root hair growth in socsld2 mutants. This indicates that SoCSLD2 was successfully knocked out in spinach root hairs using the CRISPR/Cas9-based gene editing system.
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