靶向编辑水稻Wxb基因启动子以降低稻米直链淀粉含量
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湖南省自然科学基金项目(2025JJ50110);湖南省重点研发计划项目(2023NK2004)


Targeted editing of the rice Wxb gene promoter to reduce amylose content in grains
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    摘要:

    利用CRISPR/Cas9基因编辑技术靶向编辑粳稻Wxb基因核心启动子区,通过微调Wxb基因表达水平,降低稻米直链淀粉含量。从Wxb基因启动子TATA box元件附近选取靶点(S7位点),构建基因编辑载体,经农杆菌介导遗传转化粳稻材料DNPB,筛选获得纯合突变体株系。采用RT-qPCR检测突变体材料中Wxb基因的转录表达水平,并结合田间试验考察其基本农艺性状,利用品质分析方法测定稻米直链淀粉含量。结果表明:与野生型相比,纯合突变体中Wxb基因的表达水平显著降低。突变体的主要农艺性状相较于野生型无显著差异。5个突变体的稻米直链淀粉含量为13.6%~16.6%,与野生型相比均显著降低。综上,靶向编辑粳稻材料Wxb基因启动子区可在不影响水稻基本农艺性状的基础上,有效降低Wxb基因表达水平,从而降低稻米直链淀粉含量。

    Abstract:

    To reduce the amylose content in rice, the core promoter region of the Wxb gene in japonica rice was targeted and edited by CRISPR/Cas9 gene editing technology to finely modulate the expression level of the Wxb gene. A target site (designated S7) near the TATA box element in the Wxb gene promoter was selected, and a CRISPR/Cas9 gene editing vector was constructed. Homozygous mutant lines were screened and obtained through Agrobacterium-mediated genetic transformation of the japonica rice cultivar DNPB. RT-qPCR was performed to detect the transcriptional expression levels of the Wxb gene, and agronomic traits as well as rice quality were evaluated. Field experiments were conducted to investigate the main agronomic traits, and quality analysis was employed to determine the amylose content of the rice grain. The result showed that compared with the wild type, the expression level of the Wxb gene in homozygous mutants was significantly reduced. No significant differences were observed in the main agronomic traits. The amylose content of rice grain in five mutant lines was 13.6%-16.6%, which was significantly lower than that of the wild type. In summary, targeted editing of the Wxb gene promoter region could effectively down-regulate the expression of the Wxb gene and subsequently reduce the amylose content of rice grain without affecting the basic agronomic traits of rice.

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吴宇贵,周逸龙,欧丽君,杨烨,封烨,马玉芬,张健,陈旺成,陈秋红*.靶向编辑水稻Wxb基因启动子以降低稻米直链淀粉含量[J].湖南农业大学学报(自然科学版),2026,52(2):13-19.

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  • 在线发布日期: 2026-07-13
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