利用CRISPR/Cas9系统定向编辑水稻SD1基因

Targeted Editing of Rice SD1 Gene Using CRISPR/Cas9 System

【目的】半矮秆水稻品种的选育和应用是水稻育种的最重大成果之一。半矮秆品种大多是半矮秆基因SD1(semi-dwarf1)功能缺失突变体,为了获得sd1突变体,本研究对SD1基因进行了定向编辑。【方法】利用CRISPR/Cas9系统,以SD1基因为靶基因,构建基因编辑载体CRISPR-SD1,用农杆菌介导的方法转化水稻恢复系申繁17和申繁24。【结果】在2个转化受体的T_0代均获得了纯合的sd1突变体,并且在T_1代株系中分离出了不含转基因序列的植株。2个品种的sd1突变体与各自的野生型相比,株高分别下降了25%左右。【结论】利用CRISPR/Cas9系统可以有效地对目的基因进行编辑,在水稻分子育种领域具有巨大的应用价值。

【Objective】In the last century, the usage of semi-dwarf rice varieties brought about significant improvements and great achievements in rice breeding. Most of the semi-dwarf rice varieties have loss-of-function mutations in the Semi-Dwarf1 （SD1） gene. In order to obtain sd1 mutant, targeted editing of SD1 gene was performed. 【Method】SD1 gene editing vector CRISPR-SD1 was constructed by using CRISPR/Cas9 system, and transformed into two rice restore lines Shenfan 17 and Shenfan 24 by the Agrobacterium-mediated method.【Result】 Homozygous sd1 mutants were obtained in T0 generation in both varieties, and vector-free sd1 mutant lines were segregated from the T1 population. The plant heights of the two sd1 mutants were decreased by 25% as compared with those of their wild types. 【Conclusion】CRISPR/Cas9 is a powerful tool for rice target gene editing, and has enormous potential in rice molecular breeding.