| 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. |
| 关键词: 水稻 CRISPR/Cas9 Wx基因 直链淀粉含量（AC） |
| DOI：10.20192/j.cnki.JSHNU(NS).2024.04.007 |
| 分类号:Q344 |
| 基金项目: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） |
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| Creation of Waxy allelic mutants of the rice Waxy gene using CRISPR/Cas9 technology |
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ZHU Mengke1, WANG Wenwen1, CHEN Yu1, SUN Hongfei1, CAO Liming2, LIN Dongzhi1, DONG Yanjun1
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1.College of Life Sciences, Shanghai Normal University, Shanghai 200234, China;2.Crop Breeding and Cultivation Research Institute, Shanghai Academy of Agricultural Sciences, Shanghai 201403, China
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| Abstract: |
| 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. |
| Key words: rice CRISPR/Cas9 Wx gene amylose content（AC） |
