Scientia Agricultura Sinica ›› 2025, Vol. 58 ›› Issue (8): 1650-1662.doi: 10.3864/j.issn.0578-1752.2025.08.015

• RESEARCH NOTES • Previous Articles    

Analysis of VvBES1-1 Involvement in Flower Bud Differentiation of Red Globe Grape Based on Red and Blue Light Regulation

TANG XueShen(), DANG ShiZhuo, ZHOU Juan, LI JiaHao, LI MeiHua, HU Hao, ZHANG YaHong()   

  1. School of Enology and Horticulture, Ningxia University, Yinchuan 750021
  • Received:2024-09-09 Accepted:2024-12-23 Online:2025-04-16 Published:2025-04-21
  • Contact: ZHANG YaHong

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Abstract:

【Objective】 BRI1-EMS-Suppressor 1 (BES1), a key transcription factor in brassinosteroid (BR) signaling, regulates plant photomorphogenesis and photoperiodic flowering. This study aimed to investigate the role of BES1 in flower bud differentiation of Vitis vinifera Red Globe under red and blue light regulation, thereby elucidating the BR-mediated mechanisms driven by light quality and providing insights into flowering regulation in other woody fruit trees.【Method】 Bioinformatics analysis was performed to characterize the protein structure and sequence alignment of VvBES1-1. Flower buds of Red Globe grape were collected under greenhouse natural light (CK, control) and red:blue=4:1 (R4B1) light treatment. qPCR was used to analyze the spatiotemporal expression patterns and tissue specificity of VvBES1-1 during flower bud differentiation. Subcellular localization was determined via tobacco (Nicotiana benthamiana) transient transformation. Protein-protein interactions were examined using bimolecular fluorescence complementation (BiFC), and transcriptional activation activity was assessed via yeast autoactivation assays. 【Result】VvBES1-1 contains a BES1_N domain and belongs to the BES1-S-type protein family, showing the closest phylogenetic relationship with Populus trichocarpa BES1. It is expressed throughout all developmental stages of Red Globe grapevines. During R4B1 treatment, the expression level of VvBES1-1 was significantly reduced compared to the control. Its expression peaked during the development of secondary inflorescence axes. Additionally, treatment with 10 mmol·L-¹ EBR enhanced VvBES1-1 expression in Red Globe grape flower buds. Yeast autoactivation assays demonstrated that VvBES1-1 possesses self-activation activity. Subcellular localization analysis revealed that VvBES1-1 is localized in the nucleus. Overexpression of VvBES1-1 in tobacco delayed flowering time, promoted stem elongation, and increased meristem number. Downregulation of VvBES1-1 expression integrated brassinosteroid (BR) and photoperiod signaling pathways to promote flower bud differentiation in Red Globe grapes. During flower bud differentiation, VvBES1-1 expression was significantly regulated by red and blue light. Under R4B1 treatment, its expression peaked after 6 hours of 10 mmol·L-¹ EBR exposure.【Conclusion】VvBES1-1 plays a critical role in flower bud differentiation of Red Globe grapes. It integrates BR and photoperiod signals to inhibit grape flower bud differentiation. However, under red and blue light conditions, the expression of VvBES1-1 is effectively downregulated, thereby promoting flower bud differentiation. This study provides insights into the regulatory mechanisms of light and phytohormones in grape reproductive development.

Key words: Red Globe grape, VvBES1-1, red and blue light, BR, bud differentiation

Table 1

The relevant primers required for this study"

功能
Function		 引物名称
Primer name		 引物序列
Primer sequence (5′-3′)
编码区（CDS）扩增
Coding region (CDS) amplification		 VvBES1-1-1-pCR-F ATGACAGGAACGAGGCTCCC
VvBES1-1-pCR-R TCTGGTTCTAGAGCTCCCAAGG
亚细胞定位载体构建
Subcellular localization vector construction		 VvBES1-221-F ATGACAGGAACGAGGCTCCCAACATG
VvBES1-221-R TCTGGTTCTAGAGCTCCCAAGG
酵母自激活载体构建
Yeast self-activating vector construction		 VvBES1-BD-F ATGACAGGAACGAGGCTCCC
VvBES1-BD-R TCTGGTTCTAGAGCTCCCAAGG
葡萄内参基因
Internal reference genes of grape		 VvActin-F GATTCTGGTGATGGTGTGAGT
VvActin-R GACAATTTCCCGTTCAGCAGT
VvBES1表达模式分析（葡萄）
VvBES1 expression pattern (Vitis vinifera)		 VvBES1-1-F TGGACGGTTGAAGAAGACGG
VvBES1-1-R TGGGCTTGGCTGGTATGATG
烟草荧光定量内参基因
Internal reference genes of tobacco		 NT-Actin-F GGCTTACATTGCTCTTGACTATGAAC
NT-Actin-R ATCAGGCAGCTCTGTAGCTCTCTCT
VvBES1表达模式分析（烟草）
VvBES1 Expression Pattern (Tobacco)		 VvBES1-1-NT-F GCTAGTGCTGATGCCAATGC
VvBES1-1-NT-R GTGGGGGAACTTGGATGAGG

Table 2

Physicochemical properties of VvBES1-1 protein in Red Globe grape"

蛋白质
Protein		 开放阅读框
Reading frame
(bp)		 氨基酸数
Amino acids number		 分子量
Molecular weight (kDa)		 等电点
pI		 蛋白质疏水性
Grand average of hydropathy		 亚细胞定位预测
Subcellular localization prediction
VvBES1-1 1026 341 3.629628 8.53 -0.611 细胞核Nucleus

Fig. 1

Bioinformatics analysis of the VvBES1-1 protein in Red Globe grape A: Secondary structure; B: NLS sequence prediction; C: Tertiary structure"

Fig. 2

A rootless phylogenetic tree of grape VvBES1-1 and 5 other plant BES1 proteins"

Fig. 3

Sequence comparison of VVBES1-1 protein with BES1 protein in other plant species Red boxes highlight the BES1-N structural domains"

Fig. 4

Analysis of VvBES1-1 expression levels in Red Globe grape A: Analysis of VvBES1-1 expression under different light qualities at various grape flower bud stages; B: Expression levels of VvBES1-1 in different tissues; C: Expression analysis of VvBES1-1 under different EBR (Epibrassinolide) concentrations. Different lowercase letters indicate significant differences at P<0.05. The same as below"

Fig. 5

Subcellular localization of VvBES1-1 protein in the Red Globe grape"

Fig. 6

Auto-activation activity analysis of VvBES1-1n the Red Globe grape"

Fig. 7

BiFC analysis of VvBES1-1 and VvFT"

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