高羊茅生物钟假应答因子FaPRR95的克隆、亚细胞定位及表达分析

Cloning,Subcellular Localization and Expression Analysis of Pseudo-Response Regulator 95 Genes(FaPRR95)in Tall Fescue

生物钟调节植物的生长和发育,以响应周围环境变化。假应答因子家族Pseudo-Response Regulator(PRR)是生物钟的组成部分之一,能控制光周期开花反应,家族成员表达受昼夜节律控制。为研究生物钟假应答因子PRR95基因在高羊茅中的昼夜表达模式,采用cDNA末端快速扩增技术从高羊茅叶片中克隆PRR95基因并命名为FaPRR95。该基因编码区全长为1890 bp,含1854 bp的开放阅读框,共编码617个氨基酸,定位于细胞核。系统进化树分析显示,FaPRR95蛋白与禾本科山羊草、小麦的亲缘关系最近。荧光定量PCR表明,FaPRR95基因在长日照与短日照下均有明显的昼夜节律,即使在连续光照/黑暗条件下,短期内还能维持一定的昼夜节律。

Circadian clocks regulate plant growth and development of plant in response to environmental changes.The Pseudo-response regulator(PRR)gene family is an integral component of the circadian clock,controlling photoperiodic flowering responses and exhibiting expression patterns controlled by diurnal rhythms.To investigate the diurnal expression pattern of the PRR95 gene in tall fescue,the cDNA rapid amplification of end sequences(cDNA-RACE)technique was employed to clone the PRR95 gene from tall fescue leaves,subsequently designated as FaPRR95.The full coding sequence of FaPRR95 was 1890 bp,featuring an open reading frame of 1854 bp encoding 617 amino acids and localizing within the cell nucleus.Phylogenetic analysis showed that the FaPRR95 protein had the closest evolutionary relationship to Gramineae plants such as Aegilops tauschii and Tritium aestivum.Fluorescence quantitative PCR indicated significant diurnal rhythms of FaPRR95 gene expression under both long-day and short-day conditions,maintain a certain diurnal rhythm even under continuous light/dark conditions.