VvRF2b interacts with VvTOR and influences VvTOR-regulated sugar metabolism in grape

IF 4.2 2区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Plant Science Pub Date : 2024-10-01 DOI:10.1016/j.plantsci.2024.112276

Shuang Xia , Ying Zhao , Qiaoyun Deng , Xiaoyu Han , Xiuqin Wang

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引用次数: 0

Abstract

The production of top-quality wines is closely related to the quality of the wine grapes. In wine grapes (Vitis vinifera L., Vv), sugar is a crucial determinant of berry quality, regulated by an interplay of various transcription factors and key kinases. Many transcription factors involved in sugar metabolism remain unexplored. Target of Rapamycin (TOR) is an important protein kinase in plants, recently found to regulate sugar metabolism in grapes. However, transcription factors or other factors involved in this process are rarely reported. Here, we utilized transgenic callus tissues from 'Cabernet Sauvignon' grape fruit engineered via gene overexpression (oe) and CRISPR/Cas9-based gene knockout (ko), and discovered a bZIP transcription factor, VvRF2b, whose knockout resulted in increased accumulation of fructose and sucrose, indicating that VvRF2b is a negative regulator of sugar accumulation. Subcellular localization and transcriptional activation tests showed that VvRF2b is an activator of transcription located both in the nucleus and cell membrane. Analysis of VvRF2b and VvTOR gene levels and sugar contents (glucose, fructose, and sucrose) in 'Cabernet Sauvignon' grape fruits at 30, 70, and 90 days after bloom (DAB) revealed that VvRF2b is expressed more highly during fruit development, while VvTOR is expressed more during the sugar accumulation phase, furthermore, VvTOR gene levels in koVvRF2b transgenic calli increased significantly, suggesting a strong relationship between the knockout of VvRF2b and the overexpression of VvTOR. Additionally, bimolecular fluorescence complementation and luciferase complementation assays demonstrated the interaction between VvRF2b and VvTOR proteins. After knocking out the VvRF2b gene in oeVvTOR calli, it was found that the knockout of VvRF2b promotes VvTOR-regulated sucrose accumulation and enhances the expression of sugar metabolism-related genes regulated by VvTOR. In summary, our results suggest that VvRF2b interacts with VvTOR protein and influences VvTOR-regulated sugar metabolism.

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VvRF2b 与 VvTOR 相互作用并影响 VvTOR 调控的葡萄糖代谢。

顶级葡萄酒的生产与酿酒葡萄的质量密切相关。在酿酒葡萄（Vitis vinifera L., Vv）中，糖分是决定浆果质量的关键因素，受各种转录因子和关键激酶的相互作用调节。许多参与糖代谢的转录因子仍未得到研究。雷帕霉素靶蛋白激酶（TOR）是植物中一种重要的蛋白激酶，最近发现它能调节葡萄的糖代谢。然而，参与这一过程的转录因子或其他因子却鲜有报道。在这里，我们利用'赤霞珠'葡萄果实的转基因胼胝体组织，通过基因过表达（oe）和基于CRISPR/Cas9的基因敲除（ko），发现了一种bZIP转录因子VvRF2b，其敲除会导致果糖和蔗糖的积累增加，表明VvRF2b是糖积累的负调控因子。亚细胞定位和转录激活测试表明，VvRF2b 是位于细胞核和细胞膜的转录激活因子。对开花后 30 天、70 天和 90 天（DAB）'赤霞珠'葡萄果实中 VvRF2b 和 VvTOR 基因水平和糖分含量（葡萄糖、果糖和蔗糖）的分析表明，VvRF2b 在果实发育过程中表达较多、此外，在 koVvRF2b 转基因胼胝体中，VvTOR 基因水平显著增加，这表明 VvRF2b 基因敲除与 VvTOR 基因过表达之间存在密切关系。此外，双分子荧光互补和荧光素酶互补实验证明了 VvRF2b 和 VvTOR 蛋白之间的相互作用。在 oeVvTOR 胼胝体中敲除 VvRF2b 基因后发现，敲除 VvRF2b 能促进 VvTOR 调控的蔗糖积累，并增强 VvTOR 调控的糖代谢相关基因的表达。总之，我们的研究结果表明，VvRF2b 与 VvTOR 蛋白相互作用，并影响 VvTOR 调控的糖代谢。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Plant Science 生物-生化与分子生物学

CiteScore

9.10

自引率

1.90%

发文量

322

审稿时长

33 days

期刊介绍： Plant Science will publish in the minimum of time, research manuscripts as well as commissioned reviews and commentaries recommended by its referees in all areas of experimental plant biology with emphasis in the broad areas of genomics, proteomics, biochemistry (including enzymology), physiology, cell biology, development, genetics, functional plant breeding, systems biology and the interaction of plants with the environment. Manuscripts for full consideration should be written concisely and essentially as a final report. The main criterion for publication is that the manuscript must contain original and significant insights that lead to a better understanding of fundamental plant biology. Papers centering on plant cell culture should be of interest to a wide audience and methods employed result in a substantial improvement over existing established techniques and approaches. Methods papers are welcome only when the technique(s) described is novel or provides a major advancement of established protocols.