CsDET2基因的鉴定及其对茶树光周期与非生物胁迫的响应分析

doi:10.13305/j.cnki.jts.2025.05.003

茶叶科学 ›› 2025, Vol. 45 ›› Issue (5): 742-756.doi: 10.13305/j.cnki.jts.2025.05.003

CsDET2基因的鉴定及其对茶树光周期与非生物胁迫的响应分析

李桂楠¹, 杨妮¹, 罗微¹, 张佳琪¹, 胡志航¹, 熊爱生², 郝建楠¹, 庄静^1,*

1.南京农业大学园艺学院,茶叶科学研究所,农业农村部华东地区园艺作物生物学与种质创制重点实验室,江苏南京 211800;
2.南京农业大学,作物遗传与种质创新利用全国重点实验室,江苏南京 211800

收稿日期:2025-03-05 修回日期:2025-07-09 出版日期:2025-10-15 发布日期:2025-10-17
通讯作者: * zhuangjing@njau.edu.cn
作者简介:李桂楠,女,硕士研究生,主要从事茶树遗传育种和分子生物学研究。
基金资助:
江苏省政策引导类计划（SZ-LYG202126）、园艺种质资源保护和创新计划（JSFEM-202212）、江苏高校优势学科建设项目（PAPD）

Identification of CsDET2 and Its Response Analysis to Photoperiod and Abiotic Stress in Camellia sinensis

LI Guinan¹, YANG Ni¹, LUO Wei¹, ZHANG Jiaqi¹, HU Zhihang¹, XIONG Aisheng², HAO Jiannan¹, ZHUANG Jing^1,*

1. Tea Research Institution, Ministry of Agriculture and Rural Affair Key Laboratory of Biology and Germplasm Enhancement of Horticultural Crops in East China, College of Horticulture, Nanjing Agricultural University, Nanjing 211800, China;
2. State Key Laboratory of Crop Genetics & Germplasm Enhancement and Utilization, Nanjing Agricultural University, Nanjing 211800, China

Received:2025-03-05 Revised:2025-07-09 Online:2025-10-15 Published:2025-10-17

摘要/Abstract

摘要： 茶树是我国重要的叶用经济作物,其生长发育受植物激素的影响。油菜素甾醇（BRs）是六大植物激素之一,参与调控茶树生长发育和逆境的响应,类固醇5α-还原酶是植物油菜素甾醇生物合成途径中的关键酶。为研究茶树中类固醇5α-还原酶的特性,从茶树‘舒茶早'中克隆获得CsDET2,分析该基因所编码蛋白的理化性质、二级与三级结构、序列特征、系统发育树、亚细胞定位,启动子顺式作用元件,并检测其在不同器官、非生物胁迫、光周期为12L/12D时的表达模式。结果显示,CsDET2的开放阅读框为783 bp,共编码260个氨基酸;CsDET2的分子量为30 158.97 Da,理论等电点为9.28,主要由α-螺旋和随机卷曲组成,含有典型的类固醇脱氢酶（Steroid dehydrogenase）结构域。与15个物种的DET2序列的多重比对结果显示,氨基酸序列的一致性达78.25%。系统发育树分析结果显示CsDET2与山梨猕猴桃AfDET2亲缘关系较近。亚细胞定位试验结果显示CsDET2定位在细胞质中。顺式作用元件分析显示,CsDET2的启动子区域包含4种光响应元件、4种逆境响应元件、3种激素响应元件以及5种转录因子结合位点。荧光定量结果显示,CsDET2在各器官中均有表达,在老叶中表达量最高;光周期为12L/12D时,CsDET2在白天的表达量先降低后升高,在夜晚的表达量先升高后降低,夜晚的表达水平整体高于白天的。CsDET2的表达受光周期调节,并响应高温与高盐等非生物胁迫,同时受外源赤霉素的调控。本研究为进一步研究茶树类固醇5α-还原酶的功能提供基础。

关键词: 茶树, 油菜素甾醇, 生物信息学分析, 光周期, 非生物胁迫

Abstract: Tea plants [Camellia sinensis (L.) O. Kuntze] is important leaf cash crops in China, and its growth and development are affected by plant hormones. As one of the six classical plant hormones, brassinosteroids (BRs) play crucial roles in regulating both developmental processes and stress adaptation in tea plants. Steroid 5α-reductase is a key enzyme in the brassinosteroid biosynthesis pathway. To characterize steroid 5α-reductase in Camellia sinensis, the CsDET2 gene was cloned from tea cultivar ‘Shuchazao'. Then, the physicochemical properties, secondary and tertiary structures, sequence characteristics, phylogenetic relationships, and subcellular localization of the encoded protein were analyzed, along with the cis-acting elements in gene promoter region. Furthermore, the expression profile of CsDET2 was examined across different organs under abiotic stress conditions and during a 12 h/12 h photoperiod cycle. The results reveal that the CsDET2 gene contains a 783 bp open reading frame encoding 260 amino acids. The CsDET2 protein has a molecular weight of 30 158.97 Da with a theoretical isoelectric point of 9.28, and is predominantly composed of α-helices and random coils, containing a characteristic steroid dehydrogenase domain. Comparative sequence alignment with DET2 proteins from 15 species shows 78.25% amino acid sequence identity. Phylogenetic analysis suggests that CsDET2 shares the closest evolutionary relationship with AfDET2 from Actinidiarufa. Subcellular localization experiments confirm that CsDET2 is localized in the cytoplasm. Analysis of cis-acting elements reveals that the promoter region of CsDET2 contains 4 types of photo-responsive elements, 4 types of stress-responsive elements, 3 types of hormone responsive elements, and 5 types of transcription factor binding sites. The fluorescence quantification results show that CsDET2 was expressed in all organs, and the expression level was the highest in the old leaves. In the 12L/12D photoperiod, the expression level of CsDET2 decreased first and then increased during the daytime, increased first and then decreased at night, and the expression level of CsDET2 at night was higher than that during the daytime. The expression of CsDET2 shows significant changes under high temperature and salt stress treatments, and was regulated by exogenous gibberellins. The results provided a theoretical basis and valuable insights for further exploring the function of tea plant steroid 5α-reductase.

Key words: Camellia sinensis, brassinosteroids, bioinformatics analysis, photoperiod, abiotic stress

中图分类号:

李桂楠, 杨妮, 罗微, 张佳琪, 胡志航, 熊爱生, 郝建楠, 庄静. CsDET2基因的鉴定及其对茶树光周期与非生物胁迫的响应分析[J]. 茶叶科学, 2025, 45(5): 742-756. doi: 10.13305/j.cnki.jts.2025.05.003.

LI Guinan, YANG Ni, LUO Wei, ZHANG Jiaqi, HU Zhihang, XIONG Aisheng, HAO Jiannan, ZHUANG Jing. Identification of CsDET2 and Its Response Analysis to Photoperiod and Abiotic Stress in Camellia sinensis[J]. Journal of Tea Science, 2025, 45(5): 742-756. doi: 10.13305/j.cnki.jts.2025.05.003.

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CsDET2基因的鉴定及其对茶树光周期与非生物胁迫的响应分析

Identification of CsDET2 and Its Response Analysis to Photoperiod and Abiotic Stress in Camellia sinensis

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