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青稞HvERF039基因的克隆及功能研究

马娟娥，姚有华，姚晓华，吴昆仑*，崔永梅*   

  1. 青海大学农林科学院 / 青海大学省部共建三江源生态与高原农牧业国家重点实验室 / 青藏高原种质资源研究与利用实验室 / 青海省青稞遗传育种重点实验室, 青海西宁 810016
  • 收稿日期:2025-01-24 修回日期:2025-06-01 接受日期:2025-06-01 网络出版日期:2025-06-19
  • 基金资助:
    本研究由省部共建三江源生态与高原农牧业国家重点实验室自主研究项目(2023-ZZ-01), 青海大学青年科研基金项目(2022-QNY-3), 青海省农林科学院创新基金项目(2022-NKY-04)和青藏高原种质资源研究与利用实验室项目(2025)资助。

Cloning and functional analysis of HvERF039 gene in Qingke (Hordeum vulgare L. var. nudum Hook. f)

MA Juan-E, YAO You-Hua, YAO Xiao-Hua, WU Kun-Lun*, and CUI Yong-Mei*   

  1. Academy of Agriculture and Forestry Sciences, Qinghai University / State Key Laboratory of Plateau Ecology and Agriculture, Qinghai University / Laboratory for Research and Utilization of Qinghai Tibet Plateau Germplasm Resources / Qinghai Key Laboratory of Hulless Barley Genetics and Breeding, Xining 810016, Qinghai, China
  • Received:2025-01-24 Revised:2025-06-01 Accepted:2025-06-01 Published online:2025-06-19
  • Supported by:
    This study was supported by the Open Project of State Key Laboratory of Plateau Ecology and Agriculture (2023-ZZ-01), the Qinghai University Natural Science Foundation for Young Scholars (2022-QNY-3), the Innovation Fund of Qinghai Academy of Agricultural and Forestry Sciences (2022-NKY-04), and the Laboratory for Research and Utilization of Qinghai Tibet Plateau Germplasm Resources (2025). 

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摘要：

AP2/ERF (APETALA2/ethylene responsive factor)转录因子在植物非生物胁迫响应中起重要作用，但青稞中该类基因的功能研究较少。本研究克隆了青稞HvERF039基因，并通过生物信息学、qRT-PCR、拟南芥异源过表达等方法研究了其在低温胁迫中的功能。生物信息学分析结果显示，HvERF039是具有典型AP2保守结构域的亲水性不稳定蛋白，其启动子区域含有与光响应、植物激素响应、低温胁迫响应等相关的顺式作用元件。亚细胞定位和转录自激活活性试验表明，HvERF039是一个定位于质膜和细胞核上，且具有转录激活活性的转录因子。qRT-PCR分析表明，HvERF039在低温胁迫下显著上调表达，其在不同组织中均有表达，且在叶中的表达量最高。低温胁迫下的功能验证结果显示，拟南芥转基因HvERF039过表达株系的萌发率和存活率显著高于野生型，且抗逆生理指标离子渗漏率、H2O2含量、MDA含量和CAT活性优于野生型。此外，HvERF039与多个逆境胁迫相关蛋白之间存在物理互作关系。本研究表明，HvERF039在青稞低温胁迫响应中的具有正调控作用，为青稞耐低温品种改良提供了新的基因资源。

关键词: 青稞, HvERF039, 转录因子, 异源表达, 低温胁迫

Abstract:

AP2/ERF (APETALA2/ethylene responsive factor) transcription factors play crucial roles in responses to abiotic stresses, but their functions remain poorly understood in Qingke (hulless barley). In this study, we cloned the HvERF039 gene and investigated its role in cold stress response through bioinformatics analysis, qRT-PCR, and heterologous expression in Arabidopsis thaliana. HvERF039 encoded a hydrophilic, unstable protein containing a typical AP2 conserved domain. The promoter region of HvERF039 contained various cis-acting elements, including those responsive to light, hormones and cold stress. Subcellular localization and transactivation assays revealed that HvERF039 protein was localized in both the membrane and nucleus and possessed transcriptional activation activity. qRT-PCR analysis showed that HvERF039 expression was significantly upregulated under cold treatment, with the highest expression observed in leaves and detectable levels in most other tissues. Functional analysis demonstrated that transgenic Arabidopsis plants overexpressing HvERF039 exhibited significantly higher germination and survival rates under cold stress compared to wild-type plants. These transgenic lines also showed enhanced physiological resistance, including lower ion leakage, reduced H2O2 and MDA contents, and increased CAT activity. Furthermore, HvERF039 was found to physically interact with multiple stress-related proteins. Collectively, these findings suggest that HvERF039 plays a positive regulatory role in cold stress tolerance and represents a promising genetic resource for improving cold resistance in Qingke cultivars.

Key words: Qingke, HvERF039, transcription factors, heterologous overexpression, cold stress

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