作物学报2018，Vol.44Issue(6)：844-851,8.DOI:10.3724/SP.J.1006.2018.00844
基于CRISPR/Cas9技术的水稻温敏不育基因tms5突变体的构建
Construction of tms5 Mutants in Rice Based on CRISPR/Cas9 Technology
摘要
Abstract
In order to create excellent temperature sensitive sterile line and promote the development of two line hybrid rice breeding. a 20 bp guide RNA (gRNA) was targeted to the first exon of TMS5, a thermo-sensitive genic male sterile gene as the editable object in present study, and driven by the rice U3 promoter. The target site of gRNA was ligated into the vector pCAMBIA1301, and its transgenic lines were obtained via Agrobacterium-mediated transformation. The genomic DNA of T0 transgenic lines was then extracted, and the TMS5 locus was detected by PCR and sequenced. The mutagenesis frequency for TMS5 was 63.89%, of which the frequency of homozygous deletion mutation was 34.78%. Furthermore, the fertility transition temperature and agronomic traits of T1 homozygous deletion mutants were investigated, showing that the fertility transition temperature of tms5 mutants was 28°C. Both seed setting rate and grain yields in tms5 mutants were significantly decreased compared with those in wild-type plants in paddy field condition. Together, the successful construction of tms5 mutants lays a material foundation for breeding the thermo-sensitive male sterile line of japonica rice.关键词
水稻/基因编辑/CRISPR/Cas9/TMS5/温敏不育Key words
rice/genome editing/CRISPR/Cas9/TMS5/thermo-sensitive genic male sterile引用本文复制引用
黄忠明,符辰建,杨远柱,周延彪,唐晓丹,赵新辉,周在为,符星学,王凯,史江伟,李艳锋..基于CRISPR/Cas9技术的水稻温敏不育基因tms5突变体的构建[J].作物学报,2018,44(6):844-851,8.基金项目
本研究由国家转基因生物新品种培育重大专项(2016ZX08001-004)和湖南省重点研发计划项目(2016JC2026)资助. This study was supported by the Major Project of China on New Varieties of GMO Cultivation (2016ZX08001-004) and the Key Research Program of Hunan Province (2016JC2026). （2016ZX08001-004）
