利用CRISPR/Cas9技术创制Rc基因恢复红稻

【目的】将栽培稻品种恢复为米质优、抗逆性强的红稻具有较大的研究价值。利用CRISPR/Cas9基因编辑技术，编辑原花青素转录调节因子Rc基因，恢复红种皮特性，以改良水稻米质，提升抗逆性。【方法】利用CRISPR/Cas9技术，以Rc为靶基因，构建突变载体p YLCRISPR/Cas9-Rc-g RNA，以空育180、上育453为材料，转化获得转基因植株，通过测序手段和表型观察验证成果。【结果】分子水平检测获得Rc突变材料2种，其中KY-1在1414―1417bp缺失4个碱基，终止子突变为苯丙氨酸；SY-1在1411bp处缺失1个碱基，终止子突变为天冬氨酸。2种编辑材料均恢复为红米表型，且具有一定耐盐碱能力。【结论】利用CRISPR/Cas9基因编辑技术成功获得恢复红种皮表型的纯合株系，为红米改良提供基础材料。

Breeding of Rc Function Restoration Red Rice via CRISPR/Cas9 Mediated Genome Editing

【Objective】 It is of great value to restore cultivated rice varieties to red rice with good rice quality and strong stress resistance. CRISPR/Cas9 gene editing technology was used to edit the Rc gene, restoring the red seed coat, and laying a data foundation for the improvement of rice quality and resistance.【Methods】 Using CRISPR/Cas9 technology, the target gene Rc mutant vector pYLCRISPR/ Cas9-Rc-gRNA was constructed and transformed into transgenic plants with Kongyu 180 and Shangyu 453 as materials. The results were verified by sequencing and phenotypic observation. 【Results】 Two Rc mutants were obtained. For KY-1, with 4 bases deleted from 1414 bp to 1417 bp, the terminator was replaced by phenylalanine. SY-1 also lost a base at 1411 bp, which resulted in the transformation from the terminator to aspartic acid. The two kinds of editing materials restored red rice phenotype and had certain saline-alkali tolerance.【Conclusion】 Using CRISPR/Cas9 technique, we obtained homozygous lines with red seed coat, providing basic materials for the improvement of red rice.