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

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doi：10.3969/j.issn.1000-4440.2026.04.001

关键词：水稻；4-羟基苯丙酮酸双加氧酶（HPPD）；引导编辑；flag 标签

中图分类号：S511

文献标识码：A

文章编号：1000-4440（2026）04-0649-10

Generation of rice germplasm with in situ flag tag insertion at HPPD gene mediated by prime editing

GUO Dongshu， LI Xiao， JIN Yaru， YANG Yuwen

（Institute of Germplasm Resources and Biotechnology， Jiangsu Academy of Agricultural Sciences， Nanjing ， China）

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 timepoints 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.

Key words： rice; 4-hydroxyphenylpyruvate dioxygenase （HPPD）; prime editing; flag tag

4-羟基苯丙酮酸双加氧酶（HPPD）是酪氨酸代谢途径的关键酶，可催化4-羟基苯丙酮酸（HPPA）生成尿黑酸（HGA），在植物体内，HGA是合成质体醌和生育酚的前体物质 [1-2] 。（剩余17642字）