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利用CRISPR/Cas9技术创制水稻Waxy基因等位突变体

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

riceCRISPR/Cas9Wx geneamylose content(AC)

朱梦珂、王雯雯、陈瑜、孙宏飞、曹黎明、林冬枝、董彦君

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上海师范大学生命科学学院,上海 200234

上海市农业科学院作物育种与栽培研究所,上海 201403

水稻 CRISPR/Cas9 Wx基因 直链淀粉含量(AC)

2024

上海师范大学学报(自然科学版)
上海师范大学

上海师范大学学报(自然科学版)

影响因子:0.255
ISSN:1000-5137
年,卷(期):2024.53(4)