中国水稻科学 ›› 2022, Vol. 36 ›› Issue (6): 572-578.DOI: 10.16819/j.1001-7216.2022.211205

• 研究报告 • 上一篇    下一篇

利用CRISPR/Cas9技术创制Rc基因恢复红稻

张元野1, 尹丽颖1, 李荣田1,*(), 何明良2, 刘欣欣3, 潘婷婷4, 田晓杰2, 卜庆云2, 李秀峰2,*()   

  1. 1.黑龙江大学 生命科学学院, 哈尔滨 150080
    2.中国科学院 东北地理与农业生态研究所, 哈尔滨 150081
    3.东北林业大学 生命科学学院,哈尔滨 150040
    4.黑龙江八一农垦大学 农学院, 黑龙江 大庆 163319
  • 收稿日期:2021-12-07 修回日期:2022-03-10 出版日期:2022-11-10 发布日期:2022-11-10
  • 通讯作者: 李荣田,李秀峰
  • 基金资助:
    黑龙江省杰出青年基金资助项目(JQ2020C003);中国科学院战略性先导科技专项(A类)(XDA28100000)

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

ZHANG Yuanye1, YIN Liying1, LI Rongtian1,*(), HE Mingliang2, LIU Xinxin3, PAN Tingting4, TIAN Xiaojie2, BU Qingyun2, LI Xiufeng2,*()   

  1. 1. College of Life Science, Heilongjiang University, Harbin 150080, China
    2. Northeast Institute of Geography and Agroecology, Key Laboratory of Soybean Molecular Design Breeding, Chinese Academy of Sciences, Harbin 150081, China
    3. College of Life Science, Northeast Forestry University, Harbin 150040, China
    4. College of Agriculture, Heilongjiang Bayi Agricultural University, Daqing 163319, China
  • Received:2021-12-07 Revised:2022-03-10 Online:2022-11-10 Published:2022-11-10
  • Contact: LI Rongtian, LI Xiufeng

摘要:

【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pYLCRISPR/Cas9-Rc-gRNA,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―1417 bp缺失4个碱基,终止子突变为苯丙氨酸;SY-1在1411 bp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。

关键词: 水稻, Rc, 红种皮, 耐盐碱, CRISPR/Cas9

Abstract:

【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.

Key words: rice, Rc, red episperm, salinity tolerance, CRISPR/Cas9