茶树STOP基因家族的鉴定及表达模式分析

龙露; 汤丹丹; 陈玮; 谭礼强; 陈盛相; 唐茜

doi:10.13305/j.cnki.jts.2024.03.001

茶叶科学 >

2024 , Vol. 44 >Issue 3: 386 - 398

DOI: https://doi.org/10.13305/j.cnki.jts.2024.03.001

研究报告

茶树STOP基因家族的鉴定及表达模式分析

龙露 ,
汤丹丹 ,
陈玮 ,
谭礼强 ,
陈盛相 ,
唐茜

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1.四川农业大学园艺学院,四川成都 611130;
2.精制川茶四川省重点实验室,四川成都 611130

龙露,女,硕士研究生,主要从事茶树栽培与遗传育种研究。

收稿日期: 2023-11-10

修回日期: 2024-04-09

网络出版日期: 2024-07-08

基金资助

国家自然科学基金（32202538）、四川省自然科学基金（2022NSFSC0180、2023NSFSC0166）、四川省科技厅重点研发项目（2021YFN0004）、雅安市科技计划项目（22SXHZ0057）、特色茶树新品种高效繁育技术集成与示范（2022YSX12）、雅安名山省级农业科技园区现代茶产业关键技术集成研究与示范（2021YFN0031）

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Identification and Expression Pattern Analysis of STOP Gene Family in Tea Plants (Camellia sinensis)

LONG Lu ,
TANG Dandan ,
CHEN Wei ,
TAN Liqiang ,
CHEN Shengxiang ,
TANG Qian

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1. College of Horticulture, Sichuan Agricultural University, Chengdu 611130, China;
2. Tea Refining and Innovation Key Laboratory of Sichuan Province, Chengdu 611130, China

Received date: 2023-11-10

Revised date: 2024-04-09

Online published: 2024-07-08

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摘要

STOP（Sensitive to proton rhizotoxicity）是一类C₂H₂型锌指结构转录因子,在植物多种胁迫耐受机制中发挥重要调控作用。基于茶树（Camellia sinensis）全基因组数据共鉴定出6个STOP家族基因,并运用生物信息学和实时荧光定量PCR（qRT-PCR）等方法对其进行分析。结果表明,6个CsSTOPs基因编码376~505个氨基酸,蛋白质分子量为42.17~56.36 kDa,理论等电点为5.53~8.85,均为不稳定蛋白;蛋白质保守结构域分析发现,它们均含zf-C₂H₂保守结构域;系统进化分析显示,茶树的STOP基因与拟南芥、甜橙、烟草的同源性较高;启动子顺式作用元件分析发现,CsSTOPs具有许多与生长发育、激素响应及非生物胁迫相关的作用元件;茶树各器官的转录组数据分析结果表明,CsSTOP1在根、果实、成熟叶片中的表达量较高,CsSTOP2在幼嫩叶片中的表达量较高,CsSTOP3在老叶中的表达量较高,而CsSTOP4和CsSTOP5在各个器官中的表达都较低。CsSTOPs基因能够被PEG诱导的干旱胁迫、盐胁迫、茉莉酸甲酯胁迫和冷胁迫处理诱导表达,说明CsSTOPs基因参与调控茶树生长发育和响应非生物胁迫过程。qRT-PCR检测发现,CsSTOPs、CsGS1s和CsGDHs基因在高NH₄⁺浓度处理（4.5 mmol·L^-1）的峨眉问春茶树叶和根中的表达量均高于对照处理（CK）,尤其是叶中CsSTOPs、CsGS1.1、CsGS1.3和CsGDH2在高NH₄⁺浓度处理下的表达量显著高于对照。研究结果初步解析了CsSTOPs的基本特征和功能,发现CsSTOPs可响应高NH₄⁺浓度处理,可能与CsGS1s和CsGDHs协同调控茶树适应高NH₄⁺环境的过程。

关键词： STOP基因家族; 茶树; 基因表达; 生物信息学分析

本文引用格式

龙露 , 汤丹丹 , 陈玮 , 谭礼强 , 陈盛相 , 唐茜 . 茶树STOP基因家族的鉴定及表达模式分析[J]. 茶叶科学, 2024 , 44(3) : 386 -398 . DOI: 10.13305/j.cnki.jts.2024.03.001

Abstract

STOP (Sensitive to proton rhizotoxicity) is a type of C₂H₂ zinc finger transcription factor, and it plays an important regulatory role in various stress tolerance mechanisms in higher plants. A total of 6 STOP genes were identified based on the whole genome data of tea plant (Camellia sinensis), and analyzed by bioinformatics and real-time fluorescence quantitative PCR. The results show that the six CsSTOP genes encoded 376-505 amino acids, their molecular weights were 42.17-56.36 kDa, and their theoretical isoelectric points were 5.53-8.85, all of which were unstable proteins. Conserved domain analysis of the proteins shows that they all contained zf-C₂H₂ conserved domain. Phylogenetic analysis shows that tea plant has high homology with Arabidopsis, Citrus sinensis and Nicotiana tabacum. Cis-acting element analysis of the promoter shows that CsSTOPs contain many elements related to growth and development, hormone response and abiotic stress. Transcriptome data analysis of different tissues shows that the expression level of CsSTOP1 was the higher in roots, fruits and mature leaves, the expression level of CsSTOP2 was the higher in young leaves, the expression level of CsSTOP3 was the higher in old leaves, and the expression levels of CsSTOP4 and CsSTOP5 were low in all tissues. The expressions of different CsSTOP genes were induced by PEG-induced drought stress, salt stress, methyl jasmonate stress and cold stress, indicating that CsSTOP genes were involved in the regulation of growth and development of tea plants and response to abiotic stress. Fluorescence quantitative PCR detection shows that the expression levels of CsSTOPs, CsGS1s and CsGDHs in leaves and roots of 'Emeiwenchun' treated with high NH₄⁺ concentration (4.5 mmol·L^-1) were higher than those in the control treatment (CK). Particularly, the expression levels of CsSTOPs, CsGS1.1, CsGS1.3 and CsGDH2 were significantly higher than CK in leaves treated with high NH₄⁺ concentration. In this study, the basic characteristics and functions of CsSTOPs were preliminarily analyzed, and it was found that CsSTOPs could coordinate with CsGS1s and CsGDHs genes to regulate the process of tea plant adaptation to high NH₄⁺ environmental availability.

Key words： STOP gene family; bioinformatics analysis; gene expression; tea plant

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