【摘要】 [目的]本文旨在克隆菊花ERA1(ENHANCED RESPONSE TO ABA)基因，并分析其表达模式，为探究其在调控菊花开花中的作用奠定基础。[方法]从夏菊品种‘优香’的转录组数据库中筛选得到菊花中编码法尼基转移酶β亚基基因，并命名为CmERA1。提取野生型菊花‘神马’叶片总RNA,以反转录后的cDNA为模板，对该基因cDNA全长进行克隆。运用生物信息学方法对其亲缘关系及其编码蛋白的特性进行分析预测；利用酵母单杂交系统对其转录激活活性进行分析；以绿色荧光蛋白GFP作为标签确定其亚细胞定位情况；利用实时荧光定量PCR对其在不同组织及不同光周期下的表达水平进行分析。[结果]CmERA1基因开放阅读框(ORF)全长1 356 bp,编码451个氨基酸，相对分子质量为50.0×10~3,理论pI为5.12。系统进化分析显示其与青蒿中ERA1亲缘关系最近。蛋白质二级结构分析表明，CmERA1蛋白中α-螺旋含量最高。转录激活活性分析表明，CmERA1蛋白没有转录激活活性；亚细胞定位表明，CmERA1定位于细胞核中。CmERA1基因启动子分析表明，其含有分生组织表达元件、光响应元件、ABA和SA响应元件。荧光定量PCR结果表明，CmERA1在营养生长时期的叶片中表达最高，根中表达最低；在生殖生长时期的根中表达最高，舌状花中表达最低。CmERA1的表达能够响应光周期而呈节律性的变化。[结论]CmERA1基因的表达模式分析表明其响应光周期变化。更多还原

【Abstract】 [Objectives]This article aimded to clone the ERA1(ENHANCED RESPONSE TO ABA)gene of Chrysanthemum morifolium,and analyze its expression pattern preliminarily, so as to lay the foundation for exploring its role in regulating the flowering of chrysanthemums. [Methods]The gene encoding farnesyltransferase β subunit in chrysanthemum was screened from the transcriptome database of the summer chrysanthemum ‘Yuuka’,and designated as CmERA1. The total RNA from the leaves of wild-type C.morifolium‘Jinba’was extracted, and the full-length cDNA of CmERA1 was cloned using the total cDNA of‘Jinba’leaves as a template. Bioinformatics methods were applied to analyze the genetic relationship of CmERA1 and characteristics of its encoded proteins, and yeast one-hybrid system was applied to analyze transcriptional activity of CmERA1. Real-time fluorescent quantitative PCR(RT-qPCR)was used to analyze their expression levels in different tissues and under different photoperiods. Green fluorescent protein GFP was applied as a label to determine subcellular location of CmERA1. [Results]CmERA1 gene had 1 356 bp open reading frame, encoding 451 amino acids. Its molecular mass was 50.0×10~3,and its theoretical pI was 5.12. Hylogenetic analysis showed that it had the closest relationship with ERA1 in Artemisia annua. The secondary structure analysis of CmERA1 showed that the percentage of the alpha helix was the biggest. The analysis of transcriptional activation activity showed that CmERA1 protein had no transcriptional activation activity. Subcellular localization analysis showed that CmERA1 was localized in the nucleus. The analysis of the promoter of CmERA1 showed that it contained elements related to the response to the meristem expression, light, ABA and SA. The RT-qPCR showed that the expression level was the highest in leaves during vegetative growth, the lowest in roots, the highest in roots during reproductive growth, and the lowest in ray florets. CmERA1 changed rhythmically in response to photoperiod. [Conclusions]The analysis of expression pattern showed that CmERA1 responded to photoperiod changes.更多还原