利用CRISPR/Cas 9基因编辑技术提高贵州禾产量研究

Study on Yield Improvement of‘Guizhouhe’Based on CRISPR/Cas 9 Gene Editing Technology

以贵州禾糯稻黎平杂边禾为研究材料,设计水稻产量控制关键基因OsCKX2的CRISPR/Cas 9编辑特异性靶点序列,构建水稻OsCKX2基因的CRISPR/Cas 9定点编辑载体,并利用农杆菌介导法导入黎平杂边禾,通过潮霉素筛选及PCR分子检测,获得20个独立的转基因植株。对筛选获得的转基因T0代植株基因进行测序比对,获得7株OsCKX2基因靶位点被编辑的突变转基因植株。对纯合的突变体T1代植株农艺性状进行分析发现,与野生型黎平杂边禾相比,突变体的穗粒数增加16.67%,单株产量增加25.03%。本研究获得了产量增加的黎平杂边禾基因编辑的新种质。

In this study,the Lipingzabianhe,a glutinous rice variety of Guizhouhe,was selected as materials to designed CRISPR/Cas 9-editing specific target sequences of rice yield-related gene OsCKX2,and CRISPR/Cas 9 site-specific editing vector for OsCKX2 gene was also constructed.Simultaneously,the Agrobacterium-mediated method was used to introduce target sequence into Lipingzabianhe,and 20 independent transgenic rice plants were obtained based on hygromycin screening and PCR determination.The sequencing was performed on the T0 generation of transgenic plants and 7 transgenic plants with OsCKX2 mutation were obtained.The agronomic traits of the T1-generation homozygous mutants were analyzed,and the results showed the number of grains per spike and yield per plant of the mutant were respectively increased by 16.67%and 25.03%compared with the wild type.To sum up,a new gene-editing germplasm of Lipingzabianhe with increased yield was obtained.