[1]郭冬姝,李笑,靳亚茹,等.引导编辑介导的水稻HPPD基因原位插入flag标签编码序列的遗传材料创制[J].江苏农业学报,2026,42(04):649-658.[doi:doi:10.3969/j.issn.1000-4440.2026.04.001]
　GUO Dongshu,LI Xiao,JIN Yaru,et al.Generation of rice germplasm with in situ flag tag insertion at HPPD gene mediated by prime editing[J].,2026,42(04):649-658.[doi:doi:10.3969/j.issn.1000-4440.2026.04.001]
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引导编辑介导的水稻HPPD基因原位插入flag标签编码序列的遗传材料创制()

江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
42
期数:
2026年04期
页码:
649-658
栏目:
遗传育种·生理生化
出版日期:
2026-04-30

文章信息/Info

Title:
Generation of rice germplasm with in situ flag tag insertion at HPPD gene mediated by prime editing
作者:
郭冬姝; 李笑; 靳亚茹; 杨郁文
(江苏省农业科学院种质资源与生物技术研究所,江苏南京210014)
Author(s):
GUO Dongshu; LI Xiao; JIN Yaru; YANG Yuwen
(Institute of Germplasm Resources and Biotechnology, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China)
关键词:
水稻; 4-羟基苯丙酮酸双加氧酶(HPPD); 引导编辑; flag标签
Keywords:
rice; 4-hydroxyphenylpyruvate dioxygenase (HPPD); prime editing; flag tag
分类号:
S511
DOI:
doi:10.3969/j.issn.1000-4440.2026.04.001
文献标志码:
A
摘要:
4-羟基苯丙酮酸双加氧酶(HPPD)是生物体内酪氨酸到质体醌和生育酚合成途径中的关键酶,同时也是重要的除草剂靶标。研究水稻HPPD基因的表达调控机制对于提高水稻非生物逆境抗性和培育抗除草剂品种具有重要意义。本研究通过引导编辑(PE)技术在水稻HPPD基因的3′端精准插入3×flag标签编码序列,转基因当代鉴定到3个发生目标片段精准插入的植株。蛋白质印迹法(Western blot)检测结果表明,融合蛋白能够正常表达。自交后代的基因型鉴定结果表明,1个株系的精准插入事件遗传到了下一代,并筛选到不含转基因元件的T1代纯合精准插入3×flag标签编码序列的植株。实时定量PCR检测结果表明,在黑暗转光照后的不同时间点,水稻HPPD基因的转录水平表现出由高到低的变化趋势，而HPPD蛋白表达丰度在不同时间点没有表现出显著差异。本研究创制的HPPD蛋白融合原位flag标签的水稻种质为研究水稻HPPD基因转录和HPPD蛋白表达的调控机制提供了重要的遗传资源和理论参考。
Abstract:
4-hydroxyphenylpyruvate dioxygenase (HPPD) is a key enzyme in the synthesis pathway of tyrosine to plastoquinone and tocopherol in organisms, and it is also an important target for herbicide. Investigation of transcriptional and translational regulatory mechanisms of HPPD gene is of great significance for improving rice abiotic stress resistance and developing herbicide-resistant rice varieties. In this study, the coding sequence of 3×flag tag was precisely inserted into the 3′ end of rice HPPD gene through prime editing (PE) technology. Three T0 generation plants with precise insertion of the coding sequence of 3×flag tag were identified. Western blot results indicated that the fusion protein of HPPD-3×flag expressed normally. The genotyping results of self-pollinated T1 generation plants indicated that the precise insertion event of one T0 generation plant was inherited to the next generation, and T1 generation homozygous plants harbouring precise insertion without transgenic elements were identified. The results of real-time quantitative PCR revealed that at different time points after the transition from dark to light, the transcriptional level of rice HPPD gene exhibited a trend from high to low, while the expression abundance of HPPD protein was comparable at different time points. The rice germplasm expressing HPPD-3×flag fusion protein in situ generated in this research provides an important genetic material for investigating the regulatory mechanisms of rice HPPD gene at translational and post-translational levels.

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备注/Memo

备注/Memo:
收稿日期：2025-11-25基金项目：江苏省重点研发计划现代农业项目(BE2022365)作者简介：郭冬姝(1988-),女,辽宁锦州人,博士,副研究员,主要从事水稻基因编辑技术和抗除草剂育种研究。(E-mail)guodongshuguodong@163.com
更新日期/Last Update: 2026-05-11