PsRGL1 negatively regulates chilling- and gibberellin-induced dormancy release by PsF-box1-mediated targeting for proteolytic degradation in tree peony.

IF 7.6 Q1 GENETICS & HEREDITY 园艺研究(英文) Pub Date : 2023-03-13 eCollection Date: 2023-05-01 DOI:10.1093/hr/uhad044
Linqiang Gao, Demei Niu, Tianyu Chi, Yanchao Yuan, Chunying Liu, Shupeng Gai, Yuxi Zhang
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引用次数: 2

Abstract

Tree peony bud endodormancy is a common survival strategy similar to many perennial woody plants in winter, and the activation of the GA signaling pathway is the key to breaking endodormancy. GA signal transduction is involved in many physiological processes. Although the GA-GID1-DELLA regulatory module is conserved in many plants, it has a set of specific components that add complexity to the GA response mechanism. DELLA proteins are key switches in GA signaling. Therefore, there is an urgent need to identify the key DELLA proteins involved in tree peony bud dormancy release. In this study, the prolonged chilling increased the content of endogenously active gibberellins. PsRGL1 among three DELLA proteins was significantly downregulated during chilling- and exogenous GA3-induced bud dormancy release by cell-free degradation assay, and a high level of polyubiquitination was detected. Silencing PsRGL1 accelerated bud dormancy release by increasing the expression of the genes associated with dormancy release, including PsCYCD, PsEBB1, PsEBB3, PsBG6, and PsBG9. Three F-box protein family members responded to chilling and GA3 treatments, resulting in PsF-box1 induction. Yeast two-hybrid and BiFC assays indicated that only PsF-box1 could bind to PsRGL1, and the binding site was in the C-terminal domain. PsF-box1 overexpression promoted dormancy release and upregulated the expression of the dormancy-related genes. In addition, yeast two-hybrid and pull-down assays showed that PsF-box1 also interacted with PsSKP1 to form an E3 ubiquitin ligase. These findings enriched the molecular mechanism of the GA signaling pathway during dormancy release, and enhanced the understanding of tree peony bud endodormancy.

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PsRGL1通过PsF-box1介导的牡丹蛋白水解降解靶向,负调控低温和赤霉素诱导的休眠释放。
牡丹芽内胚层是一种常见的冬季生存策略,与许多多年生木本植物相似,GA信号通路的激活是打破内胚层的关键。GA信号转导涉及许多生理过程。尽管GA-GID1-DELLA调控模块在许多植物中是保守的,但它有一组特定的成分,这增加了GA反应机制的复杂性。DELLA蛋白是GA信号传导的关键开关。因此,迫切需要鉴定参与牡丹芽休眠释放的关键DELLA蛋白。在本研究中,长期冷藏增加了内源活性赤霉素的含量。通过无细胞降解测定,三种DELLA蛋白中的PsRGL1在低温和外源GA3诱导的芽休眠释放过程中显著下调,并检测到高水平的多泛素化。沉默PsRGL1通过增加与休眠释放相关的基因(包括PsCYCD、PsEBB1、PsEBB3、PsBG6和PsBG9)的表达来加速芽休眠释放。三个F-box蛋白家族成员对冷藏和GA3处理有反应,导致PsF-box1诱导。酵母双杂交和BiFC分析表明,只有PsF-box1可以与PsRGL1结合,并且结合位点位于C末端结构域。PsF-box1过表达促进休眠释放并上调休眠相关基因的表达。此外,酵母双杂交和下拉分析表明,PsF-box1也与PsSKP1相互作用,形成E3泛素连接酶。这些发现丰富了GA信号通路在休眠释放过程中的分子机制,增强了对牡丹芽内胚层的理解。
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