Journal of Nuclear Agricultural Sciences ›› 2022, Vol. 36 ›› Issue (2): 302-312.DOI: 10.11869/j.issn.100-8551.2022.02.0302

• Induced Mutations for Plant Breeding·Agricultural Biotechnology • Previous Articles     Next Articles

Cloning, Differential Expression and Subcellular Localization of FeTOC1 Gene in Tall Fescue

WANG Qian1(), WU Jiahai2, CHEN Ying1, WANG Xiaoli1,*()   

  1. 1Guizhou Institute of Prataculture, Guizhou Academy of Agricultural Sciences, Guiyang, Guizhou 550006
    2Guizhou Institute of Fruit Tree Science, Guizhou Academy of Agricultural Sciences, Guiyang, Guizhou 550006
  • Received:2020-11-18 Accepted:2021-01-07 Online:2022-02-10 Published:2022-01-17
  • Contact: WANG Xiaoli


王茜1(), 吴佳海2, 陈莹1, 王小利1,*()   

  1. 1贵州省农业科学院草业研究所,贵州 贵阳 550006
    2贵州省农业科学院果树研究所,贵州 贵阳 550006
  • 通讯作者: 王小利
  • 作者简介:王茜,女,主要从事牧草逆境生理及分子生物学研究。E-mail:
  • 基金资助:


TOC1 gene is a major component of the central oscillator, which regulates the response to illumination through photoperiod pathway in Arabidopsis thaliana. In order to illustrate the biological function of FeTOC1 in Festuca elata, we obtained the full-length cDNA sequence of FeTOC1 by homologous cloning, analyzed the expression patterns of FeTOC1 under diverse light levels, and conducted the subcellular localization of FeTOC1. The results indicated that the full-length of FeTOC1 was 1 557 bp, encoding 517 amino acid residues; the ended protein FeTOC1 contained the pseudo-receiver domain of about 120 amino acids at N-terminus and the short CCT motif of about 50 amino acids at C-terminus; the phylogenetic tree analysis showed that FeTOC1 had close relations to those in barley, wheat and coarse goat grass, which were up to 99%. Then, the constructed target protein was fused with the GFP expression vector, and injected into tobacco, the expression suggested that FeTOC1 was mainly located in the nucleus. Real-time fluorescence quantitative analysis of the expression patterns of FeTOC1 in leaves revealed that the expression of FeTOC1 had a diurnal rhythm under both long- and short-day, and the expression at illumination 0~4 h reached the highest; compared with continuous darkness, continuous illumination weakened the oscillatory rhythm of FeTOC1 dramatically; reversing the circadian rhythm, FeTOC1 was preferentially regulated by environmental signal. To sum up, FeTOC1 gene played an important role in circadian clock regulation in tall fescue.

Key words: Festuca elata (tall fescue), TOC1 gene, expression pattern, subcellular localization


拟南芥TOC1基因是中央振荡器的重要组成部分,其编码的蛋白质TOC1通过光周期途径调控拟南芥对光照的响应。为了揭示高羊茅FeTOC1基因的生物学功能,本研究通过同源克隆获得该基因全长cDNA序列,对其在不同光照水平下的表达模式进行分析,并对其编码的蛋白质进行亚细胞定位。结果显示,高羊茅FeTOC1基因的全长cDNA序列为1 557 bp,编码517个氨基酸; 其编码的蛋白质N端存在大约120个氨基酸组成的非典型PR结构域,C端则存在大约50个氨基酸构成的CCT结构域; 系统进化树分析显示高羊茅FeTOC1蛋白质与禾本科大麦、小麦及粗山羊草亲缘关系高达99%。将构建的目的蛋白与GFP融合表达载体注射入烟草中,发现FeTOC1主要定位在细胞核中。实时荧光定量分析发现,叶片中FeTOC1表达在长、短日照下表现出一定的昼夜节律性,表达峰值均出现在光照0~4 h;相比于持续黑暗,持续光照显著地削弱了FeTOC1表达的振荡节律;颠倒昼夜节律,FeTOC1基因的表达首先受外界昼夜交替信号的调节。综上所述,FeTOC1基因在高羊茅生物钟调节中发挥着重要作用。本研究结果为进一步探索FeTOC1基因的功能,进而培育适应短日照地区牧草新品种提供了一定的理论基础。

关键词: 高羊茅, TOC1基因, 表达模式, 亚细胞定位