CsGPDH在茶籽油脂积累中的功能分析

doi:10.3864/j.issn.0578-1752.2026.02.013

摘要/Abstract

摘要：

【目的】 油脂含量是衡量油料作物品质和经济价值的关键指标。茶树（Camellia sinensis (L.) O. Kuntze）作为一种具有重要经济价值的植物，其果实茶籽在油脂资源开发领域具有潜在的应用价值。甘油-3-磷酸脱氢酶基因（GPDH）作为植物油脂合成代谢途径中的重要调控基因，克隆CsGPDH并对其表达特性与功能进行深入剖析，旨为解析CsGPDH调控茶树油脂合成的分子机制提供理论依据。【方法】 以‘金茶18’为材料，克隆CsGPDH的全长序列，运用生物信息学手段分析其编码氨基酸序列的结构特征及进化关系。采用实时荧光定量PCR技术（qRT-PCR）检测CsGPDH在茶树不同组织及油脂合成关键时期的表达模式，并通过激光共聚焦显微镜观察其编码蛋白的亚细胞定位。通过农杆菌介导的遗传转化技术，获得过表达拟南芥植株，对转基因拟南芥植株和野生型植株进行表型分析，测定种子油脂含量、脂肪酸组成等指标。【结果】 成功克隆获得CsGPDH，其编码区全长1 128 bp，CsGPDH编码的蛋白具有典型的甘油-3-磷酸脱氢酶结构域，与其他物种的同源蛋白具有较高的序列相似性。亚细胞定位结果显示，CsGPDH蛋白定位在细胞膜和细胞质。qRT-PCR分析发现，CsGPDH在种子发育的中后期表达量显著升高，与油脂快速积累期高度吻合。转基因拟南芥功能验证表明，过表达CsGPDH可使种子油脂含量提高20.6%—25.2%，不饱和脂肪酸（C18:1、C18:2）比例显著增加。【结论】 CsGPDH受种子发育进程调控，在油脂合成关键时期高表达，过量表达CsGPDH能够显著提高植株种子的油脂含量，在植物油脂合成代谢过程中发挥着重要的正向调控作用。

关键词: 茶树, 茶籽, 甘油-3-磷酸脱氢酶基因, CsGPDH, 油脂含量, 脂肪酸组分

Abstract:

【Objective】 Oil content is a key indicator determining the quality and economic value of oil crops. Tea plant (Camellia sinensis (L.) O. Kuntze), as an economically significant plant, holds potential application value in oil resource development through its fruit, tea seeds. The glycerol-3-phosphate dehydrogenase gene (GPDH) is a crucial regulatory gene in plant oil biosynthesis. Cloning CsGPDH and conducting an in-depth analysis of its expression characteristics and function aims to provide a theoretical basis for elucidating the molecular mechanism by which CsGPDH regulates oil synthesis in tea plants. 【Method】 Using ‘Jincha No.18’ as plant material, the full-length sequence of CsGPDH was cloned. Bioinformatics tools were employed to analyze the structural characteristics and evolutionary relationships of its encoded amino acid sequence. Real-time quantitative PCR (qRT-PCR) was used to detect the expression pattern of CsGPDH in different tea plant tissues and during key stages of oil synthesis. Subcellular localization of the encoded protein was observed using laser scanning confocal microscopy. Agrobacterium-mediated genetic transformation was utilized to generate CsGPDH-overexpressing Arabidopsis thaliana plants. Phenotypic analysis was performed on transgenic and wild-type Arabidopsis plants, measuring indicators such as seed oil content and fatty acid composition. 【Result】 CsGPDH was successfully cloned, with a coding sequence (CDS) length of 1 128 bp. The protein encoded by CsGPDH possesses a typical glycerol-3-phosphate dehydrogenase domain and exhibits high sequence similarity with homologous proteins from other species. Subcellular localization revealed that the CsGPDH protein is localized to the plasma membrane and cytoplasm. qRT-PCR analysis showed that CsGPDH expression significantly increased during the middle and late stages of seed development, highly coinciding with the period of rapid oil accumulation. Functional validation in transgenic Arabidopsis demonstrated that overexpression of CsGPDH increased seed oil content by 20.6%-25.2% and significantly elevated the proportion of unsaturated fatty acids (C18:1, C18:2). 【Conclusion】 CsGPDH expression is regulated by the seed developmental process, showing high expression during the critical phase of oil synthesis. Overexpression of CsGPDH can significantly enhance seed oil content in plants and plays an important positive regulatory role in the plant oil biosynthesis metabolic pathway.

Key words: Camellia sinensis, tea seeds, glycerol-3-phosphate dehydrogenase gene, CsGPDH, oil content, fatty acid composition

沈英姿, 李朵姣, 江丽, 胡新荣, 陈斌, 郑寨生, 袁名安. CsGPDH在茶籽油脂积累中的功能分析[J]. 中国农业科学, 2026, 59(2): 402-412.

SHEN YingZi, LI DuoJiao, JIANG Li, HU XingRong, CHEN Bin, ZHENG ZhaiSheng, YUAN MingAn. Functional Analysis of the CsGPDH Gene in Seed Oil Accumulation of Tea Plant[J]. Scientia Agricultura Sinica, 2026, 59(2): 402-412.

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引物名称Primers name	引物序列Primers sequence (5′-3′)	用途Function
CsGPDH-F	CTCTCTCTCTCTCTCTCTCTCTCTCTC	ORF分离 ORF isolated
CsGPDH-R	GTTTGGTACCGAAATTTACATAAA	ORF分离 ORF isolated
qRT-CsGPDH-F	AAACAGGAAATGTGCGGA	qRT-PCR
qRT-CsGPDH-R	ACAATGGCTGATGGTGGA	qRT-PCR
Actin-F	GCCATCTTTGATTGGAATGG	茶树内参基因 Tea plant endogenous control
Actin-R	GGTGCCACAACCTTGATCTT	茶树内参基因 Tea plant endogenous control
PR101-CsGPDH-F	TTGATACATATGCCCGTCGACATGGCTCCATTCTTGGAAT	亚细胞定位、转基因表达载体 Subcellular localization, transgenic expression vector
PR101-CsGPDH-R	GCTCACCATGGATCCGGTACCGTAAAATTGGGCAGCGCTT