中国水稻科学 ›› 2020, Vol. 34 ›› Issue (5): 406-412.DOI: 10.16819/j.1001-7216.2020.0104

• 研究报告 • 上一篇    下一篇

利用CRISPR/Cas9技术高效创制长粒香型水稻

徐善斌1, 郑洪亮1, 刘利锋3, 卜庆云2, 李秀峰2,*(), 邹德堂1,*()   

  1. 1东北农业大学 寒地粮食作物种质创新与生理生态教育部重点实验室, 哈尔滨 150030
    2中国科学院东北地理与农业生态研究所, 哈尔滨 150081
    3黑龙江省种业技术服务中心, 哈尔滨 150008
  • 收稿日期:2020-01-10 修回日期:2020-04-16 出版日期:2020-09-10 发布日期:2020-09-10
  • 通讯作者: 李秀峰,邹德堂
  • 基金资助:
    转基因生物新品种培育专项(2018ZX0800102B);国家重点研发专项(2017YFD0100501);黑龙江省自然科学基金(面上项目)(C2018065);国家自然科学基金青年基因资助项目(31801327);黑龙江科技厅重点研发项目(GA18B101)

Improvement of Grain Shape and Fragrance by Using CRISPR/Cas9 System

Shanbin XU1, Hongliang ZHENG1, Lifeng LIU3, Qingyun BU2, Xiufeng Li2,*(), Detang ZOU1,*()   

  1. 1Key Laboratory of Germplasm Enhancement, Physiology and Ecology of Food Crops in Cold Region, Ministry of Education, Northeast Agricultural University, Harbin 150030, China
    2Northeast Institute of Geography and Agroecology, Key Laboratory of Soybean Molecular Design Breeding, Chinese Academy of Sciences, Harbin 150081, China
    3Heilongjiang Seed Industry Technical Service Center, Harbin 150008
  • Received:2020-01-10 Revised:2020-04-16 Online:2020-09-10 Published:2020-09-10
  • Contact: Xiufeng Li, Detang ZOU

摘要:

【目的】CRISPR/Cas9基因编辑技术已成为水稻分子育种的重要手段。为了促进水稻育种的发展,本研究以非香型粳稻品种龙粳11为试验材料,对GS3GS9Badh2基因进行编辑,以期获得能稳定遗传的长粒香水稻材料。【方法】利用CRISPR/Cas9技术,以GS3GS9Badh2为靶基因,构建敲除载体pYLCRISPR/Cas9-GS3/ GS9/Badh2-gRNA,通过农杆菌介导法,在龙粳11的GS3GS9Badh2基因中引入了特定的突变。【结果】T2代无转基因的gs3/gs9/badh2纯合突变体与野生型龙粳11相比,粒长增加26.43%~27.01%,单株产量增加10.82%~12.11%,千粒重增加18.34%~41.36%,稻米变香,高效地将圆粒水稻变成长粒香型水稻。【结论】利用CRISPR/Cas9技术获得能够稳定遗传并具有长粒香品质的纯合突变株系,为组合多个品质性状提供了一种方便有效的方法,从育种角度加快了新品系创制过程。

关键词: 水稻, 粒型, 品质育种, 香米, CRISPR/Cas9

Abstract:

【Objective】 CRISPR/Cas9-mediated genome editing has become an important way for molecular breeding in rice. To promote rice breeding, the non-fragrant japonica rice variety Longjing11 was used as the test material. This study aims to edit GS3, GS9 and Badh2 genes for obtaining valuable and stable long-grain fragrant rice materials. 【Method】 The target genes GS3, GS9 and BADH2 were selected to construct the vector pYLCRISPR/ Cas9-GS3/ GS9/Badh2-gRNA by using CRISPR/Cas9 gene editing technology. The Agrobacterium-mediated transformation was used to transform Longjing11. Specific mutations were introduced into the GS3, GS9 and Badh2 genes in Longjing 11. 【Result】 The grain length of the transgenic free homozygote gs3/gs9/badh2 increased by 26.43% to 27.01%, yield per plant by 10.82% to 12.11%, 1000-grain weight by 18.34% to 41.36%, rice became fragrant as compared with wild type of Longjing 11. This study efficiently transformed round-grain rice into long-grain fragrant rice. 【Conclusion】The homozygous mutant lines featured by stable inheritance and long-grain fragrant quality were obtained by using CRISPR/Cas9 system. This study provides a convenient and effective way of combining multiple quality traits together, which could significantly accelerate breeding process from a breeding perspective.

Key words: rice, grain shape, quality breeding, fragrant rice, CRISPR/Cas9

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