甘薯IbSAUR76基因表达特征及互作蛋白筛选
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1.山西农业大学生命科学学院;2.山西农业大学农学院;3.山西农业大学棉花所;4.山西农业大学玉米所;5.山西农业大学高粱研究所

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山西省研究生科研创新项目(2023KY321);中央引导地方科技发展资金项目(YDZJSX2024B007);山西农业大学校科技创新提升工程(CXGC2023051;CXGC2023074);山西省回国留学人员科研资助项目(2021-074);山西省基础研究计划(202403021212098);山西农业大学杂粮研究院项目(Z120220502)


Identification of IbSAUR76 Gene Expression Profiles and Screening of Interacting Proteins in Sweetpotato
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The Graduate Research and Innovation Projects of Shanxi Province (2023KY321); The Central Guidance for Local Science and Technology Development Fund Project (YDZJSX2024B007); Science and Technology Innovation Enhancement Programs of Shanxi Agricultural University (CXGC2023051; CXGC2023074); Research Funding Project for Returned Overseas Students in Shanxi Province (2021-074); Shanxi Province Basic Research Program (202403021212098); Projects of the Millet Research Institute of Shanxi Agricultural University (Z120220502)

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    摘要:

    甘薯[Ipomoea batatas (L.) Lam.]在整个生长发育过程中容易遭受各种各样的生物胁迫和非生物胁迫,这些胁迫严重影响其生长发育,进而导致其品质下降和产量锐减。因此,挖掘甘薯抗逆相关基因并深入探究其功能与作用机制,进而培育抗逆性强的甘薯新品种对于提升甘薯的耐盐耐旱性、保障甘薯产业的可持续发展具有重要意义。SAUR(small auxin up RNA)基因作为生长素信号传导途径中的关键成员,在调控植物生长发育和响应非生物胁迫方面发挥着重要作用。为了深入探究IbSAUR76基因在甘薯中的功能及其作用机制,本研究成功克隆了IbSAUR76基因,并构建了35::IbSAUR76::GFP融合表达载体,亚细胞定位结果显示IbSAUR76蛋白定位于细胞核。为了进一步揭示IbSAUR76基因在甘薯逆境胁迫中的作用机制,我们克隆了IbSAUR76基因的启动子序列。顺式作用元件预测结果表明,IbSAUR76中包含多个逆境和激素响应元件。在干旱和盐处理条件下,IbSAUR76的表达量均显著升高,表明该基因在甘薯的逆境应答中发挥作用。此外,本研究构建了IbSAUR76的诱饵载体,通过mating法从甘薯cDNA酵母文库中筛选得到27个与IbSAUR76相互作用的候选蛋白,并对其中两个与非生物胁迫相关的互作蛋白IbERF2及IbHOS15进行酵母双杂交回转验证。本研究丰富了对IbSAUR76基因功能的认识,为深入解析IbSAUR76在甘薯中的功能和作用机制提供了理论基础。

    Abstract:

    Sweetpotato [Ipomoea batatas (L.) Lam.] is highly susceptible to a variety of biotic and abiotic stresses during its growth and development. These stresses can severely disrupt its growth and development, which in turn causes a significant decline in quality and yield. Therefore, identifying stress-resistance-related genes in sweetpotato, exploring their functions and mechanisms, and breeding new varieties with enhanced stress resistance are crucial for improving salt and drought tolerance and ensuring the sustainable development of the sweetpotato. SAUR (small auxin up RNA) genes are key components of the auxin signaling pathway and play essential roles in regulating plant growth and development as well as in responding to abiotic stresses. To investigate the function and mechanism of the IbSAUR76 gene in sweetpotato, we cloned this gene and constructed an IbSAUR76-GFP fusion expression vector. Our results showed that the IbSAUR76 protein is localized in the nucleus. To gain deeper insights into the role of the IbSAUR76 gene in the stress response of sweetpotato, we then cloned its promoter sequence. The prediction of cis-acting elements revealed that the IbSAUR76 promoter contains multiple stress and hormone response elements. Under drought and salt treatments, the expression level of IbSAUR76 was significantly upregulated, indicating that this gene plays a role in the stress response of sweetpotato. Moreover, a bait vector for IbSAUR76 was constructed and used to screen the sweetpotato cDNA yeast library, identifying 27 candidate proteins that interact with IbSAUR76. Among these, two interaction proteins related to abiotic stress, IbERF2 and IbHOS15, were verified using yeast two-hybrid back-transformation. In summary, this study not only enriches our understanding of the IbSAUR76 gene in sweetpotato but also provides a solid theoretical foundation for further in-depth analysis of its function. The identification and characterization of IbSAUR76 and its interacting proteins are significant steps toward developing more stress-resistant sweetpotato varieties. Among these, two interaction proteins related to abiotic stress, IbERF2 and IbHOS15, were verified by yeast two-hybrid (Y2H). In summary, this study not only enriches our knowledge of the IbSAUR76 gene in sweetpotato but also provides a theoretical basis for further in-depth analysis of its function.

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  • 收稿日期:2024-11-13
  • 最后修改日期:2025-01-18
  • 录用日期:2025-01-21
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