State Key Laboratory of Plant Genomics, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, China.
Plant J. 2014 Dec;80(6):1118-30. doi: 10.1111/tpj.12714.
Seed germination is a key developmental process in the plant life cycle that is influenced by various environmental cues and phytohormones through gene expression and a series of metabolism pathways. In the present study, we investigated a C2C2-type finger protein, OsLOL1, which promotes gibberellin (GA) biosynthesis and affects seed germination in Oryza sativa (rice). We used OsLOL1 antisense and sense transgenic lines to explore OsLOL1 functions. Seed germination timing in antisense plants was restored to wild type when exogenous GA3 was applied. The reduced expression of the GA biosynthesis gene OsKO2 and the accumulation of ent-kaurene were observed during germination in antisense plants. Based on yeast two-hybrid and firefly luciferase complementation analyses, OsLOL1 interacted with the basic leucine zipper protein OsbZIP58. The results from electrophoretic mobility shift and dual-luciferase reporter assays showed that OsbZIP58 binds the G-box cis-element of the OsKO2 promoter and activates LUC reporter gene expression, and that interaction between OsLOL1 and OsbZIP58 activates OsKO2 gene expression. In addition, OsLOL1 decreased SOD1 gene expression and accelerated programmed cell death (PCD) in the aleurone layer of rice grains. These findings demonstrate that the interaction between OsLOL1 and OsbZIP58 influences GA biosynthesis through the activation of OsKO2 via OsbZIP58, thereby stimulating aleurone PCD and seed germination.
种子萌发是植物生命周期中的一个关键发育过程,受到各种环境线索和植物激素的影响,通过基因表达和一系列代谢途径来实现。在本研究中,我们研究了一个 C2C2 型手指蛋白 OsLOL1,它促进赤霉素(GA)的生物合成,并影响水稻中的种子萌发。我们使用 OsLOL1 反义及正义转基因系来探索 OsLOL1 的功能。在反义植物中,外源 GA3 的应用恢复了种子萌发的时间。在反义植物中,GA 生物合成基因 OsKO2 的表达减少,并且在萌发过程中积累了 ent-贝壳杉烯。基于酵母双杂交和萤火虫荧光素酶互补分析,OsLOL1 与碱性亮氨酸拉链蛋白 OsbZIP58 相互作用。电泳迁移率变动和双荧光素酶报告基因分析的结果表明,OsbZIP58 结合 OsKO2 启动子的 G 盒顺式元件,并激活 LUC 报告基因的表达,并且 OsLOL1 和 OsbZIP58 之间的相互作用激活 OsKO2 基因的表达。此外,OsLOL1 降低了 SOD1 基因的表达,并加速了水稻籽粒糊粉层中的程序性细胞死亡(PCD)。这些发现表明,OsLOL1 和 OsbZIP58 之间的相互作用通过 OsbZIP58 激活 OsKO2 来影响 GA 生物合成,从而刺激糊粉层 PCD 和种子萌发。