College of Life Sciences, Qingdao Agricultural University, Qingdao, China.
Plant J. 2020 Aug;103(4):1386-1398. doi: 10.1111/tpj.14807. Epub 2020 Jun 4.
Actin depolymerizing factor (ADF) is a key modulator for dynamic organization of actin cytoskeleton. Interestingly, it was found that the ADF1 gene silencing delays flowering, but its mechanism remains unclear. In this study, ADF1 was used as a bait to screen its interacting proteins by the yeast two-hybrid (Y2H) system. One of them, the REM16 transcription factor was identified. As one of the AP2/B3-like transcriptional factor family members, the REM16 contains two B3 domains and its transcript levels kept increasing during the floral transition stage. Overexpression of REM16 accelerates flowering while silencing of REM16 delays flowering. Gene expression analysis indicated that the key flowering activation genes such as CONSTANS (CO), FLOWERING LOCUS T (FT), LEAFY (LFY) and SUPPRESSOR OF OVEREXPRESSION OF CONSTANS (SOC1) were upregulated in the REM16 overexpression lines, while the transcription of the flowering suppression gene FLOWERING LOCUS C (FLC) was decreased. In contrast, the REM16 gene silencing lines contained lower transcript levels of the CO, FT, LFY and SOC1 but higher transcript levels of the FLC compared with the wild-type plants. It was proved that REM16 could directly bind to the promoter regions of SOC1 and FT by in vitro and in vivo assays. Genetic analysis supported that REM16 acts upstream of SOC1 and FT in flowering pathways. All these studies provided strong evidence demonstrating that REM16 promotes flowering by directly activating SOC1 and FT.
肌动蛋白解聚因子 (ADF) 是调节肌动蛋白细胞骨架动态组织的关键调节剂。有趣的是,研究发现 ADF1 基因沉默会延迟开花,但具体机制尚不清楚。在本研究中,我们以 ADF1 为诱饵,通过酵母双杂交 (Y2H) 系统筛选其相互作用蛋白。其中一个是 REM16 转录因子。作为 AP2/B3 样转录因子家族成员之一,REM16 包含两个 B3 结构域,其转录水平在花转变阶段不断增加。过表达 REM16 会加速开花,而沉默 REM16 则会延迟开花。基因表达分析表明,关键的开花激活基因,如 CONSTANS (CO)、FLOWERING LOCUS T (FT)、LEAFY (LFY) 和 SUPPRESSOR OF OVEREXPRESSION OF CONSTANS (SOC1),在 REM16 过表达系中上调,而开花抑制基因 FLOWERING LOCUS C (FLC) 的转录则下调。相比之下,REM16 基因沉默系中 CO、FT、LFY 和 SOC1 的转录水平较低,而 FLC 的转录水平较高。通过体外和体内实验证明,REM16 可以直接结合 SOC1 和 FT 启动子区域。遗传分析支持 REM16 在开花途径中位于 SOC1 和 FT 的上游。这些研究结果为 REM16 通过直接激活 SOC1 和 FT 促进开花提供了有力证据。