CRISPR/Cas9技术编辑OsRhoGDI2基因导致水稻半矮化

Disruption of OsRhoGDI2 by CRISPR/Cas9 technology leads to semi-dwarf in rice

摘要:

OsRhoGDI2是通过酵母双杂交从水稻幼穗中分离的一个与Rho蛋白家族成员OsRacD相互作用蛋白的编码基因，但功能尚不明确。本研究利用CRISPR/Cas9技术创制水稻OsRhoGDI2基因敲除突变体。检测结果表明，转基因水稻T0代获得2种纯合突变体，T1代获得8种纯合突变体。序列分析显示，在敲除水稻中，该基因的编辑靶点附近发生了碱基的替换或缺失，预期生成丧失RhoGDI保守结构域的截短蛋白。表型比对分析发现，敲除水稻与对照相比，株高显著降低，统计学分析结果显示，敲除水稻株高降低源于第Ⅱ和第Ⅲ茎节的缩短，提示OsRhoGDI2基因可能与水稻株高控制相关。

Abstract:

OsRhoGDI2 was isolated as a putative partner of Rho protein family member OsRacD from rice panicles by yeast two-hybrid, but its function remains unknown. In order to identify the function of OsRhoGDI2, OsRhoGDI2 knockout mutants were created by CRISPR/Cas9 technology. The results showed that two different homozygous mutants were obtained in T0 generation, and eight kinds homozygous mutants were identified in T1 generation. Sequence analysis revealed that the base substitution or base deletion occurred near the editing targets of the gene in knockout rice, and it could be expected that the truncated OsRhoGDI2 proteins lacking the RhoGDI conserved domain would be generated. Phenotype analysis showed that the OsRhoGDI2 knockout rice plants were significantly lower than the control plants. Statistical analysis confirmed that the significant decrease of plant height was due to the shortening of the second and third internodes, suggesting that OsRhoGDI2 gene may be related with rice height control.