长穗偃麦草EeSKOR启动子的克隆及功能分析
投稿时间:2021-07-12  修订日期:2021-09-06  点此下载全文
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张勇 北京市农林科学院草业花卉与景观生态研究所 100097
田小霞 北京市农林科学院草业花卉与景观生态研究所 100097
郑明利 北京市农林科学院草业花卉与景观生态研究所 100097
毛培春 北京市农林科学院草业花卉与景观生态研究所 100097
孟林 北京市农林科学院草业花卉与景观生态研究所 100097
基金项目:国家林业和草原局行业科技重大项目课题(LCZD202004);北京市自然科学基金项目(6182013);北京市农林科学院科技创新能力建设专项(KJCX20200107,KJCX20170110)
中文摘要:为了研究外整流钾通道蛋白(stelar K+ outward rectifier channels,SKOR)基因SKOR在长穗偃麦草中的功能,利用热不对称交错PCR(Tail-PCR)技术,克隆了长穗偃麦草EeSKOR启动子,并进行启动子顺式作用元件及基因表达分析。结果表明,EeSKOR启动子除必须具备的核心启动元件外,还含有特异转录因子结合位点、植物激素响应元件、光响应元件、组织特异的启动元件和胁迫响应元件。EeSKOR启动子驱动GUS报告基因可在拟南芥的叶、叶柄和根中表达。在NaCl、PEG、ABA和SA处理下长穗偃麦草EeSKOR基因在根中呈现不同的表达模式,NaCl处理下EeSKOR的表达量呈先下调后上调趋势;PEG处理下EeSKOR的表达量呈上调趋势,且随着时间的延长显著上调;ABA处理下EeSKOR的表达受到抑制且随处理时间延长呈显著下调趋势;SA处理下EeSKOR表现出先上调后下调趋势,且在处理72 h时表达量显著低于正常表达水平。综合分析显示,EeSKOR基因的表达受NaCl、PEG、ABA和SA的诱导调节。本研究结果为进一步系统研究长穗偃麦草EeSKOR基因功能提供重要理论依据。
中文关键词:启动子  EeSKOR  长穗偃麦草  GUS染色  非生物胁迫
 
Cloning and Functional Analysis of the EeSKOR Promoter of Elytrigia elongata
Abstract:In order to study the function of the stelar K+ outward rectifier channels (SKOR) gene in Elytrigia elongata, the EeSKOR promoter of E. elongata was cloned by the technique of thermal asymmetric cross-PCR (Tail-PCR), and the promoter cis-acting elements and gene expression analysis were performed. The results showed that the EeSKOR promoter also contains specific transcription factor binding sites, plant hormone response elements, light response elements, tissue-specific promoter elements and stress response elements in addition to the essential core promoter elements. The EeSKOR promoter drives the GUS reporter gene to be expressed in the leaves, petioles and roots of Arabidopsis. Under NaCl, PEG, ABA and SA treatments, EeSKOR showed the different expression patterns in the roots of E. elongata. The expression of EeSKOR showed a trend of first down-regulation and then up-regulation under NaCl treatment, under PEG treatment, the expression of EeSKOR gene was up-regulated, and significantly up-regulated with time, ABA treatment caused the expression of EeSKOR to be inhibited and decreased significantly with the extension of treatment time, and the expression of EeSKOR showed a trend of up-regulation and then down-regulation under SA treatment, and the expression level of EeSKOR was significantly lower than the normal expression level after 72 h of SA treatment. The comprehensive analysis showed that EeSKOR gene expression was regulated by NaCl, PEG, ABA and SA. The results of this study would provide an important theoretical basis for the further systematic study on the EeSKOR gene function in E. elongata.
keywords:Promoter  EeSKOR  Elytrigia elongata  GUS histochemical staining  Abiotic stress
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