HvNF-YB7: A Key Regulator of the HvGSTF13.1-Mediated Glutathione Antioxidant System in Drought-Tolerant Barley.

IF 3.6 2区 生物学 Q1 PLANT SCIENCES
Rui Pan, Badr Alharthi, Tiantian Wu, Lin Wang, Jingqiu Cheng, Sajid Fiaz
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Abstract

Drought stress severely constrains barley yield and quality improvement. The Nuclear Factor Y (NF-Y) transcription factor family plays a crucial role in plant stress responses; however, its biological function and molecular regulatory mechanism in barley under drought stress remain unclear. This study integrated evolutionary analysis and transcriptomics to reveal the characteristics of the barley NF-Y gene family. Molecular biology experiments were employed to elucidate the drought response function and regulatory mechanism of the core gene HvNF-YB7. Phylogenetic analysis indicated evolutionary conservation and lineage-specific expansion patterns within the NF-Y gene family. The results found that HvNF-YB7 was induced 9.63-fold by drought in the tolerant wild barley genotype EC_S1. An 8-bp deletion variant in HvNF-YB7's promoter region created a CAAT-box cis-acting element, significantly enhancing its transcriptional activity under drought stress. Functional validation using transgenic overexpression significantly improved drought tolerance (e.g., fresh weight, water content), whereas silencing by virus-induced gene silencing (VIGS) exacerbated damage. This confirmed HvNF-YB7 as a key positive regulator. The yeast two-hybrid (Y2H) and RUBY-luciferase assay revealed that HvNF-YB7 forms a transcriptional complex by interacting with NF-YA3 and then recognizes and activates the CCAAT element in the promoter of HvGSTF13.1, a key gene in the glutathione metabolic pathway. This activation enhances glutathione (GSH)-mediated reactive oxygen species (ROS) scavenging capacity, effectively mitigating drought-induced oxidative damage. Our findings elucidate the "HvNF-YB7-HvGSTF13.1-GSH" pathway, which enhances barley drought tolerance, and provide a significant theoretical foundation and genetic resources for understanding its molecular basis.

HvNF-YB7: hvgstf13.1介导的大麦谷胱甘肽抗氧化系统的关键调控因子
干旱胁迫严重制约大麦产量和品质的提高。核因子Y (NF-Y)转录因子家族在植物逆境响应中起着至关重要的作用;但其在干旱胁迫下大麦中的生物学功能和分子调控机制尚不清楚。本研究将进化分析和转录组学相结合,揭示了大麦NF-Y基因家族的特征。通过分子生物学实验,阐明核心基因HvNF-YB7的干旱响应功能和调控机制。系统发育分析表明NF-Y基因家族具有进化保守性和谱系特异性扩展模式。结果发现,HvNF-YB7在耐旱性野生大麦基因型EC_S1中被干旱诱导了9.63倍。HvNF-YB7启动子区域的一个8 bp缺失变体创建了CAAT-box顺式作用元件,显著增强了其在干旱胁迫下的转录活性。功能验证使用转基因过表达显著提高抗旱性(例如,鲜重,含水量),而通过病毒诱导的基因沉默(VIGS)沉默加剧了损害。这证实了HvNF-YB7是一个关键的阳性调节因子。酵母双杂交(Y2H)和红宝石荧光素酶实验发现,HvNF-YB7通过与NF-YA3相互作用形成转录复合体,识别并激活谷胱甘肽代谢途径关键基因HvGSTF13.1启动子中的CCAAT元件。这种激活增强了谷胱甘肽(GSH)介导的活性氧(ROS)清除能力,有效减轻了干旱引起的氧化损伤。本研究结果阐明了“HvNF-YB7-HvGSTF13.1-GSH”通路对大麦抗旱性的增强作用,为了解其分子基础提供了重要的理论基础和遗传资源。
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来源期刊
Physiologia plantarum
Physiologia plantarum 生物-植物科学
CiteScore
11.00
自引率
3.10%
发文量
224
审稿时长
3.9 months
期刊介绍: Physiologia Plantarum is an international journal committed to publishing the best full-length original research papers that advance our understanding of primary mechanisms of plant development, growth and productivity as well as plant interactions with the biotic and abiotic environment. All organisational levels of experimental plant biology – from molecular and cell biology, biochemistry and biophysics to ecophysiology and global change biology – fall within the scope of the journal. The content is distributed between 5 main subject areas supervised by Subject Editors specialised in the respective domain: (1) biochemistry and metabolism, (2) ecophysiology, stress and adaptation, (3) uptake, transport and assimilation, (4) development, growth and differentiation, (5) photobiology and photosynthesis.
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