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BEL1 样蛋白(StBEL5)通过马铃薯中串联的 TGAC 核心基序调控细胞周期蛋白 DOF 因子 1(StCDF1)。

BEL1-like protein (StBEL5) regulates CYCLING DOF FACTOR1 (StCDF1) through tandem TGAC core motifs in potato.

机构信息

Biology Division, Dr. Homi Bhabha Road, Indian Institute of Science Education and Research (IISER), Pune, 411008, Maharashtra, India.

Biology Division, Dr. Homi Bhabha Road, Indian Institute of Science Education and Research (IISER), Pune, 411008, Maharashtra, India.

出版信息

J Plant Physiol. 2019 Oct;241:153014. doi: 10.1016/j.jplph.2019.153014. Epub 2019 Jul 31.

Abstract

Tuberization in potato is governed by many intrinsic and extrinsic factors. Various molecular signals, such as red light photoreceptor (StPHYB), BEL1-like transcription factor (StBEL5), CYCLING DOF FACTOR1 (StCDF1), StCO1/2 (CONSTANS1/2) and StSP6A (Flowering Locus T orthologue), function as crucial regulators during the photoperiod-dependent tuberization pathway. StCDF1 induces tuberization by increasing StSP6A levels via StCO1/2 suppression. Although the circadian clock proteins, GIGANTEA (StGI) and FLAVIN-BINDING, KELCH REPEAT, F-BOX 1 (StFKF1), are reported as StCDF1 interactors, how the StCDF1 gene is regulated in potato is unknown. The BEL-KNOX heterodimer regulates key tuberization genes through tandem TGAC core motifs in their promoters. A recent study reported the presence of six tandem TGAC core motifs in the StCDF1 promoter, suggesting possible regulation of StCDF1 by StBEL5. In our study, we observed a positive correlation between StBEL5 and StCDF1 expression, whereas StCDF1 and its known repressor, StFKF1, showed a negative correlation for the tested tissue types. To investigate the StBEL5-StCDF1 interaction, we generated transgenic potato promoter lines containing a wild-type or mutated (deletion of six tandem TGAC sites) StCDF1 promoter fused to GUS. Wild-type promoter transgenic lines exhibited widespread GUS activity, whereas this activity was absent in the mutated promoter transgenic lines. Moreover, StBEL5 and StCDF1 transcript levels were significantly higher in the stolon-to-tuber stages under short-day conditions compared to long-day conditions. Using wild-type and mutated prStCDF1 as baits in Y1H assays, we further demonstrated that StBEL5 interacts with the StCDF1 promoter through tandem TGAC motifs, indicating direct regulation of StCDF1 by StBEL5 in potato.

摘要

块茎形成在马铃薯中受许多内在和外在因素的控制。各种分子信号,如红光光受体(StPHYB)、BEL1 样转录因子(StBEL5)、CYCLING DOF FACTOR1(StCDF1)、StCO1/2(CONSTANS1/2)和 StSP6A(Flowering Locus T 同源物),在光周期依赖性块茎形成途径中作为关键调节剂发挥作用。StCDF1 通过抑制 StCO1/2 来增加 StSP6A 水平来诱导块茎形成。虽然昼夜节律钟蛋白 GIGANTEA(StGI)和黄素结合、KELCH 重复、F-BOX 1(StFKF1)被报道为 StCDF1 的相互作用物,但马铃薯中 StCDF1 基因是如何被调控的尚不清楚。BEL-KNOX 异源二聚体通过其启动子中的串联 TGAC 核心基序来调节关键的块茎形成基因。最近的一项研究报告称,在 StCDF1 启动子中存在六个串联 TGAC 核心基序,表明 StBEL5 可能调节 StCDF1。在我们的研究中,我们观察到 StBEL5 和 StCDF1 表达之间存在正相关,而 StCDF1 和其已知的抑制剂 StFKF1 在测试的组织类型中呈负相关。为了研究 StBEL5-StCDF1 相互作用,我们生成了含有野生型或突变型(缺失六个串联 TGAC 位点)StCDF1 启动子融合到 GUS 的转基因马铃薯启动子系。野生型启动子转基因系表现出广泛的 GUS 活性,而突变型启动子转基因系则没有这种活性。此外,在短日照条件下,与长日照条件相比,匍匐茎到块茎阶段的 StBEL5 和 StCDF1 转录本水平显著升高。在 Y1H 测定中,使用野生型和突变型 prStCDF1 作为诱饵,我们进一步证明 StBEL5 通过串联 TGAC 基序与 StCDF1 启动子相互作用,表明 StBEL5 在马铃薯中直接调控 StCDF1。

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