水稻真核翻译起始因子OseIF6.2调控粒型的功能研究
收稿日期: 2024-10-25
修回日期: 2025-03-27
网络出版日期: 2025-05-21
基金资助
四川省农业科学院农业前沿技术攻关专项(5+1QYGG003);国家现代农业产业技术体系四川创新团队项目(SCCXTD-2024-1);国家重点研发计划资助项目(2024YFD1200900);国家现代农业产业技术体系建设专项(CARS-01);四川省农业科学院原始创新项目(YSCX2035-010);四川省科技计划重点研发项目(2021YFYZ0016)
Functional Study of Rice Eukaryotic Translation Initiation Factor OseIF6.2 in Grain Size Regulation
Received date: 2024-10-25
Revised date: 2025-03-27
Online published: 2025-05-21
【目的】本研究旨在通过反向遗传学策略，深入探究水稻真核翻译起始因子OseIF6.2在水稻生长发育中的具体功能，以期为OseIF6.2功能的进一步解析奠定基础。【方法】首先，利用生物信息学方法进行多重序列比对及进化树的构建，以分析OseIF6.2与其他真核生物中eIF6蛋白的系统发育关系；运用CRISPR/Cas9基因编辑技术，对水稻(日本晴)中的OseIF6.2基因进行精准编辑。其次，通过qRT-PCR技术，检测OseIF6.2在水稻不同组织中的表达水平；借助激光共聚焦显微镜，观察OseIF6.2在细胞内的精确定位。然后对野生型与oseif6.2突变体的粒长、粒宽、粒厚及千粒重等关键农艺性状进行比较分析。最后，通过酵母双杂交(Y2H)及双分子荧光互补(BiFC)实验，验证OseIF6.2与其潜在互作蛋白OseIF6.1的相互作用。【结果】系统发育分析结果表明，OseIF6.2与其他真核生物中的eIF6蛋白具有高度保守性，凸显了其在真核生物翻译调控中的普遍重要性。qRT-PCR结果显示，OseIF6.2在水稻茎中的表达水平较高。通过将OseIF6.2蛋白与GFP融合后注入本氏烟叶片细胞，并在激光共聚焦显微镜下观察，发现OseIF6.2在细胞核及细胞质区域均有荧光信号分布，明确了其亚细胞定位。此外，与野生型相比，oseif6.2突变体在粒长、粒厚及千粒重等性状上均表现为显著降低。Y2H及BiFC实验均成功验证了OseIF6.2与OseIF6.1蛋白之间的直接相互作用。【结论】综上所述，OseIF6.2作为水稻真核翻译起始因子，其功能异常显著影响水稻的粒型特征，进一步揭示了该因子在水稻生长发育及粒型形成中的关键作用。
马顺婷, 胡运高, 高方远, 刘利平, 牟昌铃, 吕建群, 苏相文, 刘松, 梁毓玉, 任光俊, 郭鸿鸣 . 水稻真核翻译起始因子OseIF6.2调控粒型的功能研究[J]. 中国水稻科学, 2025 , 39(3) : 331 -342 . DOI: 10.16819/j.1001-7216.2025.2401010
【Objective】This study aims to investigate the functional role of the rice eukaryotic translation initiation factor OseIF6.2 in rice growth and development using a reverse genetics approach, thereby laying a foundation for further exploration of the functions of OseIF6.2.【Method】First, bioinformatics methods were employed to perform multiple sequence alignment and a phylogenetic tree was constructed to analyze the evolutionary relationship between OseIF6.2 and eIF6 proteins in other eukaryotes. CRISPR/Cas9 gene editing technology was used to precisely edit the OseIF6.2 gene in rice (Nipponbare).Next, the expression levels of OseIF6.2 in different tissues of rice were detected using quantitative real-time PCR (qRT-PCR) technology. The precise localization of OseIF6.2 within cells was observed using a confocal laser microscope.Finally, the interaction between OseIF6.2 and its potential interacting protein, OseIF6.1, was validated using yeast two-hybrid (Y2H) and bimolecular fluorescence complementation (BiFC) assays. 【Result】Phylogenetic analysis revealed that OseIF6.2 is highly conserved among eukaryotic eIF6 proteins, underscoring its universal role in translation regulation.qRT-PCR results indicated that OseIF6.2 expression was relatively high in rice stems. Subcellular localization analysis using GFP-tagged OseIF6.2 in tobacco leaf cells showed fluorescence signals in both the nucleus and cytoplasm, confirming its dual localization.Additionally, compared to the wild type, the oseif6.2 mutant exhibited significant reductions in grain length, grain thickness, and 1000-grain weight. Both Y2H and BiFC experiments confirmed a direct interaction between OseIF6.2 and OseIF6.1【Conclusion】In summary, OseIF6.2, a eukaryotic translation initiation factor in rice, plays a crucial role in regulating grain morphology, further highlighting its importance in rice growth, development, and grain formation.
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