利用CRISPR/Cas9技术高效创制长粒香型水稻  

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

作　　者：徐善斌 郑洪亮[1] 刘利锋 卜庆云[2] 李秀峰[2] 邹德堂[1] XU Shanbin;ZHENG Hongliang;LIU Lifeng;BU Qingyun;Li Xiufeng;ZOU Detang(Key Laboratory of Germplasm Enhancement,Physiology and Ecology of Food Crops in Cold Region,Ministry of Education,Northeast Agricultural University,Harbin 150030,China;Northeast Institute of Geography and Agroecology,Key Laboratory of Soybean Molecular Design Breeding,Chinese Academy of Sciences,Harbin 150081,China;Heilongjiang Seed Industry Technical Service Center,Harbin 150008)

机构地区：[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

出　　处：《中国水稻科学》2020年第5期406-412,共7页Chinese Journal of Rice Science

基　　金：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)。

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GS3、GS9和Badh2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GS3、GS9和Badh2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GS3、GS9和Badh2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。【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.

关 键 词：水稻 粒型 品质育种 香米 CRISPR/Cas9 

分 类 号：Q755[生物学—分子生物学] S511.032[农业科学—作物学]