江苏农业学报2026，Vol.42Issue(4)：649-658,10.DOI:10.3969/j.issn.1000-4440.2026.04.001
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
Generation of rice germplasm with in situ flag tag insertion at HPPD gene mediated by prime editing
摘要
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 ex-pressed 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 inser-tion 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.关键词
水稻/4-羟基苯丙酮酸双加氧酶(HPPD)/引导编辑/flag标签Key words
rice/4-hydroxyphenylpyruvate dioxygenase(HPPD)/prime editing/flag tag分类
农业科技引用本文复制引用
郭冬姝,李笑,靳亚茹,杨郁文..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(4):649-658,10.基金项目
江苏省重点研发计划现代农业项目(BE2022365) （BE2022365）
