中国水稻科学2017，Vol.31Issue(3)：223-231,9.DOI:10.16819/j.1001-7216.2017.7029
利用CRISPR/Cas9系统定向改良水稻粒长和穗粒数性状
Orientation Improvement of Grain Length and Grain Number in Rice by Using CRISPR/Cas9 System
摘要
Abstract
[Objective]Gene orientation editing has become an important way for molecular breeding. We evaluated the improvement effects on the target traits following the construction of GS3 and Gn1a loss-of-function mutants so as to lay a solid foundation for high-yielding rice breeding. [[Method]GS3 and Gn1a were selected as targets for gene editing, which control the grain size and grain number in rice, respectively. The Pc1300-2×35S:: Cas9-Ggs3-gGn1a expression vector was constructed for knocking out both GS3 and Gn1a by using CRISPR/Cas9 system, and transformed into four good quality rice varieties by the Agrobacterium-mediated method. And the properties of mutations and the target traits were analyzed in the transgenic lines.[Result]The sequencing results showed the GS3 and Gn1a in four rice varieties were successfully edited. In T0 generation, we obtained mutants with frame shift mutations in GS3 and Gn1a in four rice varieties. In T1 generation, the marker-free plants were screened for analyzing the agronomic traits in four genetic backgrounds. For the agronomic characters, the gs3 and gs3gn1a mutants had the longer grain length and the increased 1000-grain weight compared to the wild type, and the gs3gn1a mutants had more grains per panicle compared to the gs3 mutants.[Conclusion]CRISPR/Cas9-mediated gene editing can improve rice target traits, which has great potential in orientation improvement of rice varieties.关键词
CRISPR/Cas9/基因编辑/水稻/GS3/Gn1aKey words
CRISPR/Cas9/gene editing/rice/GS3/Gn1a分类
生物科学引用本文复制引用
沈兰,李健,付亚萍,王俊杰,华宇峰,焦晓真,严长杰,王克剑..利用CRISPR/Cas9系统定向改良水稻粒长和穗粒数性状[J].中国水稻科学,2017,31(3):223-231,9.基金项目
国家自然科学基金资助项目(31371233) （31371233）
高校自然科学基金重大项目(14KJA210002) （14KJA210002）
中国农业科学院科技创新工程资助项目 （）
江苏省农业科技自主创新资金资助项目[CX(13)5075]. （13）
