敲除TMS5基因获得温敏不育粳稻新材料
收稿日期: 2019-03-28
修回日期: 2019-04-23
网络出版日期: 2019-09-10
基金资助
国家重点研发计划资助项目(2017YFD0100403);江苏省农业科技自主创新资金资助项目[CX(18)2022];江苏省现代农业重点研发计划资助项目(BE2017345);国家转基因生物新品种培育科技重大专项(2018ZX0800102);江苏省自然科学基金资助项目(BK20170610)
Thermo-sensitive Male Sterile Line Created by Editing TMS5 Gene in japonica Rice
Received date: 2019-03-28
Revised date: 2019-04-23
Online published: 2019-09-10
【目的】 水稻温敏不育系的选育是两系杂交稻育种工作中的关键环节。TMS5是控制温敏不育的重要基因,在生产上也应用较广。为了加快水稻温敏不育系的选育进程,【方法】 利用CRISPR/Cas9技术,在武运粳7号背景下对TMS5基因进行编辑。【结果】 通过测序从22株T0代中筛选获得6株纯合的突变体,分析发现6个纯合的突变体产生了相同的遗传变异,都在TMS5基因第2外显子44 bp和45 bp之间插入碱基“A”,导致翻译提前终止。通过细胞学观察和人工辅助授粉证实获得的株系花粉败育而雌配子正常发育。【结论】 这些结果表明对温敏不育基因TMS5基因进行编辑可以快速获得粳型温敏雄性不育系。
关键词： 粳稻; CRISPR/Cas9; 温敏雄性不育系
杜茜, 费云燕, 王芳权, 许扬, 王军, 李文奇, 赵凌, 陈智慧, 梁国华, 周勇, 杨杰 . 敲除TMS5基因获得温敏不育粳稻新材料[J]. 中国水稻科学, 2019 , 33(5) : 429 -435 . DOI: 10.16819/j.1001-7216.2019.9035
【Objective】The breeding of thermo-sensitive male sterile rice lines is a key step in the breeding of two-line hybrid rice. TMS5 is the key gene that modulates pollen fertility of rice at different temperatures and has been widely used in rice breeding. In order to accelerate the breeding process of thermo-sensitive sterility lines, 【Method】CRISPR/Cas9 technology was used to edit TMS5 with a japonica rice variety Wuyunjing 7 as a material. 【Results】Sequencing analysis showed that six homozygous mutants screened from 22 strains of T0 generation produced the same genetic variation, and one base “A” was inserted between 44 bp and 45 bp of the second exon of TMS5 of all the six mutants, resulting in pre-maturation of translation. Cytological observation and artificial pollination confirmed that the pollen of tms5 mutant was aborted and the development of female gametes was normal. 【Conclusion】These results indicated that the thermo-sensitive male sterile lines could be obtained rapidly by knocking out the thermo-sensitive male sterile gene TMS5 in japonica rice.
Key words： japonica rice; CRISPR/Cas9; thermo-sensitive male sterile line
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