甘薯IbGAPCp1在干旱和盐胁迫下的表达及上游调控因子筛选
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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.山西农业大学玉米所;4.山西农业大学高粱研究所;5.山西农业大学棉花所;6.山西农业大学玉米研究所

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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)


Expression of IbGAPCp1 in Sweetpotato under Drought and Salt stress and Screening of Upstream Regulatory Factors
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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:

    我国盐碱和干旱土地面积广大,边际土地利用率低。甘薯作为重要的块根类作物,在保障国家粮食安全方面具有重要意义。因此,挖掘甘薯重要抗逆基因并培育甘薯抗逆新品种,可有效地利用这些盐碱、干旱和边际土地,对进一步提升土地资源利用率至关重要。GAPCp1是定位于质体中的一种甘油醛-3-磷酸脱氢酶,在植物生长发育和能量代谢中起重要作用。本研究以栗子香(LZX)为材料,成功克隆了其开放阅读框(ORF)序列。亚细胞定位研究表明,IbGAPCp1编码的蛋白质定位于叶绿体中。定量PCR分析显示,IbGAPCp1在甘薯不同组织中均有表达,其中叶片中的表达量最高;随着块根的发育,IbGAPCp1基因的表达逐渐升高,在105 d时达到最高水平。为进一步探索IbGAPCp1基因的调控机制,克隆了长为1940 bp的IbGAPCp1基因启动子序列Pro-IbGAPCp1。顺式作用元件预测结果表明,Pro-IbGAPCp1中含有多个与光响应、激素响应及分生组织表达等相关的元件。干旱和盐胁迫处理下IbGAPCp1基因的表达动态结果显示,IbGAPCp1基因的表达均呈现先上升后下降的趋势,表明该基因对盐和干旱胁迫具有一定的响应。进一步通过构建诱饵载体并利用酵母单杂交技术筛选出了与IbGAPCp1启动子区相互作用的35个上游调控因子,其中包括乙烯不敏感蛋白2、蔗糖合酶2及MYB44等。这些因子与植物的生长发育、次生代谢、能量产生及多种逆境响应(如干旱、盐和低温)等相关。本研究为深入探讨IbGAPCp1在甘薯非生物胁迫中的功能及作用机制提供了理论基础。

    Abstract:

    China characterized by extensive saline-alkali and arid land, along with low utilization rate of marginal lands. As an important tuberous root crop, sweetpotato plays a significant role in ensuring national food security. Therefore, identifying important stress-resistance genes in sweetpotato and developing new resilient varieties can effectively utilize these saline-alkali, arid, and marginal lands, thereby significantly enhancing the utilization efficiency of land resources. GAPCp1, a glyceraldehyde-3-phosphate dehydrogenase located in plastid, plays a crucial role in plant growth, development, and energy metabolism. In this study, using the sweetpotato variety Lizixiang (LZX) as the material, we successfully cloned the open reading frame (ORF) sequence of IbGAPCp1. Subcellular localization studies showed that the protein encoded by IbGAPCp1 is localized in chloroplasts. Quantitative real-time PCR analysis showed that IbGAPCp1 is expressed in all tested tissues of sweetpotato, with the highest transcription level in leaf tissues. The expression of IbGAPCp1 gene increased gradually with the development of sweetpotato tuberous root and reaching its peak at 105 days. In order to further explore the regulatory mechanism of IbGAPCp1 gene, we cloned a 1940 bp sequence of the IbGAPCp1 gene promoter, designated as Pro-IbGAPCp1. The prediction of cis-acting elements showed that Pro-IbGAPCp1 contains several elements related to photoresponse, hormone response and meristem expression. Under drought and salt stress treatments, IbGAPCp1 gene exhibited a trend of first increasing and then decreasing, indicating that this gene responds to drought and salt stress. Additionally, 35 upstream regulators interacting with the promoter region of IbGAPCp1 were screened by yeast one-hybrid technique, including ethylene-insensitive protein 2, sucrose synthase 2 and MYB44. These factors are involved in plant growth and development, secondary metabolism, energy production, and various stress responses, such as drought, salt and low temperatures. This study provides a theoretical basis for further study on the function and mechanism of IbGAPCp1 in response to abiotic stress in sweetpotato.

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  • 收稿日期:2024-11-13
  • 最后修改日期:2025-01-28
  • 录用日期:2025-02-05
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