CRISPR/Cas 9基因编辑技术降低贵州禾株高研究

Study on Reducing GuizhouHe Plant Height Based onCRISPR/Cas 9 Technology

为获得株高降低的贵州禾水稻新种质,以贵州禾高秆糯稻材料黎平杂边禾为研究材料,根据CRISPR/Cas 9靶位点设计原则设计水稻OsGA20ox2基因的特异性靶点sgRNA序列,构建水稻OsGA20ox2基因的CRISPR/Cas 9定点突变载体,利用农杆菌介导的愈伤转化法遗传转化黎平杂边禾,通过潮霉素筛选,以及PCR分子检测,获得16株转基因植株。对筛选获得的转基因植株基因进行测序,获得7株OsGA20ox2基因靶位点被编辑的突变转基因植株。研究表明,T 1代突变体植株株高比黎平杂边禾降低了29.6%。

In order to create the new germplasm of Guizhouhe rice with reduced plant height,the Guizhouhe rice variety Lipingzabianhe was used as research materials to design the specific target sgRNA sequence of OsGA20ox2 gene according to the design principle of CRISPR/Cas9 target sites and CRISPR/Cas 9 site-directed mutant vector of rice OsGA20ox2 gene was constructed.The results showed that 16 transgenic rice plants were obtained by using Agrobacterium-mediated callus transformation of Lipingzabianhe,hygromycin screening and PCR detection.Further,the objective gene of the screened transgenic plants were sequenced to verify the 7 mutant transgenic plants whose OsGA20ox2 gene were site-specifically edited.Additionally,the results showed that the plant height of T 1 generation mutant was 29.6%lower than that of the wild type.