小麦ERF转录因子基因TaERF1-A调控小麦耐盐性的功能研究

doi:10.3864/j.issn.0578-1752.2026.20.001

中国农业科学 ›› 2026, Vol. 59 ›› Issue (20): 4433-4446.doi: 10.3864/j.issn.0578-1752.2026.20.001

• 作物遗传育种·种质资源·分子遗传学 • 上一篇下一篇

小麦ERF转录因子基因TaERF1-A调控小麦耐盐性的功能研究

于洋¹^,³(), 李梦婷²(), 刘丹¹, 许庆芬¹, 万子语¹, 王停停³, 冯刚¹, 王建贺¹, 王从磊¹(), 梁丹¹()

¹ 天津市农业科学院农作物遗传育种重点实验室，天津 300192
² 天津农学院食品科学与生物工程学院，天津 300192
³ 西北农林科技大学/作物抗逆与高效生产全国重点实验室，陕西杨凌 712100

收稿日期:2026-02-06 接受日期:2026-04-02 出版日期:2026-10-21 发布日期:2026-10-21
通信作者:
梁丹，E-mail：liangdan188@126.com。
王丛磊，E-mail：clwang1000@126.com
联系方式: 于洋，E-mail：yuyang0319@nwafu.edu.cn。李梦婷，E-mail：lmt0802@126.com。于洋和李梦婷为同等贡献作者。
基金资助:
青年科技人才托举工程项目(TAASTJ2026-004); 西北农林科技大学作物抗逆与高效生产全国重点实验室课题基金(SKLCSRHPKF2026008); 资助技术创新引导专项(25YDTPJC00570); 天津市科技计划项目(25ZYCGSN00820)

Functional Characterization of the ERF Transcription Factor TaERF1-A in Salt Tolerance in Triticum aestivum L.

YU Yang¹^,³(), LI MengTing²(), LIU Dan¹, XU QingFen¹, WAN ZiYu¹, WANG TingTing³, FENG Gang¹, WANG JianHe¹, WANG CongLei¹(), LIANG Dan¹()

¹ Key Laboratory of Crop Genetic Breeding, Tianjin Academy of Agricultural Sciences, Tianjin 300192
² College of Food Science and Biotechnology, Tianjin Agricultural University, Tianjin 300192
³ Northwest A&F University/National Key Laboratory of Crop Stress Resistance and High-Efficiency Production, Yangling 712100, Shaanxi

Received:2026-02-06 Accepted:2026-04-02 Published:2026-10-21 Online:2026-10-21

摘要/Abstract

摘要：

【目的】ERF转录因子在调控植物非生物胁迫反应中发挥重要作用。研究小麦ERF转录因子基因TaERF1-A的耐盐功能，为小麦耐盐品种的培育提供有价值的基因资源和理论依据。【方法】使用qRT-PCR检测盐胁迫下TaERF1-A的表达模式；采用PlantCARE预测TaERF1-A启动子的顺式作用元件；利用烟草瞬时表达技术对TaERF1-A蛋白亚细胞定位进行分析；构建TaERF1-A小麦沉默载体和过表达载体，通过农杆菌介导的方法，获得小麦过表达和瞬时沉默株系；对转基因植株进行耐盐性鉴定和生理指标测定；通过酵母双杂交筛选TaERF1-A的互作蛋白，利用双分子荧光互补（BiFC）和荧光素酶互补（LCA）试验进一步验证互作关系；应用双荧光素酶（LUC）试验研究互作蛋白和TaERF1-A对胁迫相关基因启动子的影响。【结果】TaERF1-A启动子中含有多种与非生物胁迫相关的顺式调控元件，编码蛋白定位于细胞核。在盐胁迫下，TaERF1-A显著上调表达，沉默TaERF1-A小麦株系的MDA含量上升，而脯氨酸含量和抗氧化酶（POD、CAT和SOD）的活性均降低，胁迫响应（TaDREB1和TaERF3）和抗氧化酶基因（TaPOD、TaCAT和TaSOD）的表达被抑制；过表达TaERF1-A植株的MDA含量降低，而脯氨酸水平升高，POD、CAT和SOD活性增强，TaDREB1、TaERF3、TaPOD、TaCAT和TaSOD的表达水平上调，结果表明，在盐胁迫下，小麦TaERF1-A正向调控小麦耐盐性。通过酵母双杂交（Y2H）、双分子荧光互补（BiFC）和荧光素酶互补（LCA）试验，证实在酵母和烟草细胞中TaERF1-A和TaNAC48存在互作关系。TaNAC48的表达受到盐胁迫诱导，TaNAC48与TaERF1-A的共表达提高了胁迫相关基因启动子的激活活性。【结论】小麦TaERF1-A在盐胁迫过程中发挥正向调控作用，过表达和沉默TaERF1-A分别增强和减弱了转基因小麦的耐盐性；TaERF1-A可能与TaNAC48蛋白协作，调控胁迫相关基因表达，提高小麦耐盐性。

关键词: 小麦, TaERF1-A, 基因沉默，过表达, 耐盐性, 互作蛋白

Abstract:

【Objective】Ethylene response factor (ERF) transcription factors play important roles in regulating plant responses to abiotic stresses. This study aimed to investigate the function of the wheat ERF transcription factor gene TaERF1-A in salt tolerance, providing valuable genetic resources and a theoretical basis for breeding salt-tolerant wheat varieties.【Method】The expression pattern of TaERF1-A under salt stress was analyzed by qRT-PCR. Cis-acting regulatory elements in the promoter region of TaERF1-A were predicted using PlantCARE. Subcellular localization of the TaERF1-A protein was determined by transient expression in tobacco leaves. A TaERF1-A overexpression construct and a wheat gene-silencing vector were generated, and transgenic wheat lines with TaERF1-A overexpression or transient silencing were obtained via Agrobacterium-mediated transformation. Salt tolerance and related physiological indices of the transgenic plants were evaluated. Potential interacting proteins of TaERF1-A were screened using a yeast two-hybrid (Y2H) system, and the interactions were further validated by bimolecular fluorescence complementation (BiFC) and luciferase complementation assay (LCA). A dual-luciferase (LUC) assay was used to examine the effects of TaERF1-A and its interacting proteins on the promoters of stress-related genes.【Result】The TaERF1-A promoter contains various cis-regulatory elements associated with abiotic stress responses, and the encoded protein is localized in the nucleus. Under salt stress, TaERF1-A expression was significantly upregulated. Silencing of TaERF1-A in wheat resulted in increased malondialdehyde (MDA) content and decreased proline content as well as reduced activities of antioxidant enzymes (POD, CAT, and SOD) under salt stress. In addition, the expression of stress-responsive genes (TaDREB1 and TaERF3) and antioxidant enzyme genes (TaPOD, TaCAT, and TaSOD) was suppressed. In contrast, overexpression of TaERF1-A led to reduced MDA levels, increased proline accumulation, enhanced activities of POD, CAT, and SOD, and upregulated expression of TaDREB1, TaERF3, TaPOD, TaCAT, and TaSOD, indicating that TaERF1-A positively regulates salt tolerance in wheat. Y2H, BiFC, and LCA assays confirmed the interaction between TaERF1-A and TaNAC48 in yeast and tobacco cells. Moreover, the expression of TaNAC48 was induced by salt stress, and co-expression of TaNAC48 with TaERF1-A enhanced the activation of stress-responsive gene promoters. 【Conclusion】TaERF1-A acts as a positive regulator in the salt stress response of wheat. Overexpression and silencing of TaERF1-A respectively enhanced and reduced salt tolerance in transgenic wheat. TaERF1-A may cooperate with TaNAC48 to regulate the expression of stress-related genes, thereby improving salt tolerance in wheat.

Key words: wheat, TaERF1-A, gene silencing, overexpression, salt tolerance, protein interaction

于洋, 李梦婷, 刘丹, 许庆芬, 万子语, 王停停, 冯刚, 王建贺, 王从磊, 梁丹. 小麦ERF转录因子基因TaERF1-A调控小麦耐盐性的功能研究[J]. 中国农业科学, 2026, 59(20): 4433-4446.

YU Yang, LI MengTing, LIU Dan, XU QingFen, WAN ZiYu, WANG TingTing, FENG Gang, WANG JianHe, WANG CongLei, LIANG Dan. Functional Characterization of the ERF Transcription Factor TaERF1-A in Salt Tolerance in Triticum aestivum L.[J]. Scientia Agricultura Sinica, 2026, 59(20): 4433-4446.

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基因Gene	候选蛋白Putative protein	登记号Accession
TaNAC48	NAC结构域蛋白NAC domain-containing protein 48	XM_044463398.1
TaVDAC	线粒体外膜孔道蛋白Mitochondrial outer membrane porin	XM_044532654.1

单倍型 Haplotype	国内品种 Domestic cultivars	国外品种 Foreign cultivars	品种数目 Cultivar number
HapⅠ	周8425B、川麦104、济麦22、宁春4号 Zhou 8425B, Chuanmai 104, Jimai 22, Ningchun 4	Alchemy, Julius, Landmark, Mace, Stanley	9
HapⅡ	中国春、郑麦366、西农6028、矮抗58、小偃6号、北京8号、扬麦33、川麦48、扬麦158、邯6172、藏1817、济麦44、郑麦16、济麦47、郑麦22、克丰11、科农9024、石4185、苏麦3号 Chinese Spring, Zhengmai 366, Xinong 6028, Aikang 58, Xiaoyan 6, Beijing 8, Yangmai 33, Chuanmai 48, Yangmai 158, Han 6172, Zang 1817, Jimai 44, Zhengmai 16, Jimai 47, Zhengmai 22, Kefeng11, Kenong 9024, Shi 4185, Sumai 3	ArinalrFor, Claire, Jagger, Kariega, Lancer, Norin61, Robigus, Sy Mattis	27

小麦ERF转录因子基因TaERF1-A调控小麦耐盐性的功能研究

Functional Characterization of the ERF Transcription Factor TaERF1-A in Salt Tolerance in Triticum aestivum L.

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