利用CRISPR/Cas9技术创制水稻Waxy基因等位突变体  

Creation of Waxy allelic mutants of the rice Waxy gene using CRISPR/Cas9 technology

作　　者：朱梦珂 王雯雯 陈瑜 孙宏飞 曹黎明[2] 林冬枝[1] 董彦君[1] ZHU Mengke;WANG Wenwen;CHEN Yu;SUN Hongfei;CAO Liming;LIN Dongzhi;DONG Yanjun(College of Life Sciences,Shanghai Normal University,Shanghai 200234,China;Crop Breeding and Cultivation Research Institute,Shanghai Academy of Agricultural Sciences,Shanghai 201403,China)

机构地区：[1]上海师范大学生命科学学院,上海200234 [2]上海市农业科学院作物育种与栽培研究所,上海201403

出　　处：《上海师范大学学报（自然科学版中英文）》2024年第4期502-511,共10页Journal of Shanghai Normal University(Natural Sciences)

基　　金：国家自然科学基金(30971552);上海市产业技术体系建设项目[沪农科产字(2021)第3号];上海市科技兴农项目(2021-02-00-12-F00769);上海市科委项目(19391900200)。

摘　　要：利用CRISPR/Cas9基因编辑技术,在粳稻品种“嘉花1号”的颗粒结合型淀粉合成酶Ⅰ基因(Waxy,Wx)的启动子区域、5'UTR区域,以及第2、3外显子区域设计基因敲除靶点,获得4种不同类型的纯合编辑水稻突变体,随后对其进行稻米品质与农艺性状分析.结果表明:不同类型Wx纯合编辑突变体与野生型“嘉花1号”相比,核心启动子区域编辑的突变体wxpro直链淀粉含量(AC)下降至14.09%;5'UTR区域编辑的突变体wxutr的AC下降至11.76%,且淀粉粒排列松散;而在外显子区域编辑的突变体wxex2和wxex3的AC下降至2%以下,与糯稻品种类似.本研究创制了能稳定遗传且AC不同的纯合突变株系,不仅使水稻Wx基因的等位突变类型变得更加丰富,也为水稻食味品质改良提供了优良的遗传育种材料.In this study,CRISPR/Cas9 gene editing technology was used to design four knockout targets located in the promoter region,5'UTR region and exons 2 and 3 of the granule bound starch synthase gene(Waxy,Wx)in the japonica rice variety“Jiahua 1”.We then obtained four different types of T1 generation homozygous edited-mutants without transgene to investigate their quality and agronomic traits.Compared to the wild type“Jiahua 1”,the amylose content(AC)of the wxpro mutants in the promoter region decreased to 14.09%,while the wxutr mutants in the 5'UTR region decreased to 11.76%,with a loose arrangement of starch grains.The wxex2 and wxex3 mutants in the exon regions had an AC of to less than 2%,similar to that of glutinous rice.In summary,all stable-inherited homozygous wx mutants,with varying AC,not only enrich the Wx allelic mutant genes in rice,but also provide valuable genetic and breeding materials for improving rice eating quality.

关 键 词：水稻 CRISPR/Cas9 WX基因 直链淀粉含量(AC) 

分 类 号：Q344[生物学—遗传学]