利用CRISPR/Cas9技术改良南粳46抽穗期
收稿日期: 2024-07-09
修回日期: 2024-08-12
网络出版日期: 2025-11-19
基金资助
生物育种钟山实验室项目(ZSBBL-KY2023-06-3);宿迁英才群英计划青年资助项目([2022]QNXM-0035);江苏省种业振兴揭榜挂帅项目(JBGS[2021]001)
Modifying Heading Date of Nanjing 46 via CRISPR/Cas9-mediated Genome Editing
Received date: 2024-07-09
Revised date: 2024-08-12
Online published: 2025-11-19
【目的】抽穗期是决定水稻品种种植区域和生态适应性的关键农艺性状。本研究通过对优良食味中熟晚粳品种南粳46(简称NJ46)的抽穗期基因进行编辑，以缩短NJ46的生育期，扩大其种植区域。【方法】利用CRISPR/Cas9技术，以DTH8和Ghd2为靶基因，构建pCAMBIA1305-Actin:Cas9-sgRNADTH8与pCAMBIA1305-Actin:Cas9-sgRNAGhd2编辑载体，用农杆菌介导的植物转化方法转化NJ46，筛选不含外源成分的纯合编辑系，并考察其抽穗期、农艺性状及品质相关性状。【结果】对DTH8的编辑获得了3个比NJ46抽穗期提早22~36 d的无外源成分纯合突变系(NJ46-dth8-1~-3)，对Ghd2的编辑获得了3个比NJ46抽穗期提早14~15 d的无外源成分纯合突变系(NJ46-ghd2-1~-3)。分别在苏中和苏北测定了各基因纯合突变系的抽穗期、主要农艺性状及品质性状，与相应生态区推广品种南粳9108、南粳518进行比较，发现NJ46-ghd2-1单株产量显著高于南粳9108，但抽穗期和品质与之相当，好于另外2个敲除系；NJ46-dth8-1与南粳518在单株产量、主要品质性状上均无显著差异，好于另外2个敲除系。【结论】定向编辑南粳46中的DTH8和Ghd2基因，获得适宜苏北和苏中生态区种植的优良食味粳稻新种质，有助于扩大优良食味水稻品种的推广种植区域。
陈伟, 叶元妹, 赵剑华, 冯志明, 陈宗祥, 胡珂鸣, 左示敏 . 利用CRISPR/Cas9技术改良南粳46抽穗期[J]. 中国水稻科学, 2025 , 39(6) : 760 -770 . DOI: 10.16819/j.1001-7216.2025.240707
【Objective】 Heading date is a key agronomic trait determining the regional and ecological adaptation of rice cultivars. We edited the heading date genes of the medium-maturing late japonica rice variety Nanjing 46 (NJ46) with excellent tasting to shorten its heading date thereby potentially expanding its planting area.【Method】 Using CRISPR/Cas9 technology, the editing constructs, pCAMBIA1305-Actin:Cas9-sgRNADTH8 and pCAMBIA1305-Actin:Cas9-sgRNAGhd2 targeting DTH8 and Ghd2, respectively, were developed and transformed into NJ46 via Agrobacterium-mediated transformation. Homozygous editing lines without exogenous components were screened and evaluated on heading date, agronomic traits and quality-related traits. 【Result】 Three homozygous mutant lines were obtained. The NJ46-dth8 lines (NJ46-dth8-1 to -3) presented 22-36 days earlier than NJ46 on heading date, while the NJ46-ghd2 lines (NJ46-ghd2-1 to -3) showed 14-15 days earlier. The heading date, major agronomic traits, and quality related components of the homozygous mutant lines were evaluated in different areas of central and northern regions of Jiangsu Province, and compared with the widely cultivated varieties Nanjing 9108 and Nanjing 518 in their respectively ecological zones. NJ46-ghd2-1 exhibited a significantly higher yield per plant than Nanjing 9108, while showing comparable heading date and quality traits, and performed better than the other two knockout lines. NJ46-dth8-1 showed no significant differences from Nanjing 518 in yield per plant and major quality traits, and performed better than the other two knockout lines.【Conclusion】 Knockout of DTH8 and Ghd2 genes in NJ46 yielded new germplasms suitable for planting in the northern and central region of Jiangsu Province, respectively, which will help expand the planting area of this excellent-tasting rice variety.
Key words： rice; CRISPR/Cas9; genetic editing; heading date; DTH8; Ghd2
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