《植物生理学报》 2017, 53(8): 1523-1531

高羊茅FaFT2基因克隆及表达分析

陈锡^1,2, 赵德刚^1,2, 陈莹¹, 李小冬¹, 吴佳海¹, 王小利^1,2,*

¹贵州省农业科学院草业研究所, 贵阳550006; ²贵州大学喀斯特山区植物资源利用与育种国家地方联合工程研究中心, 贵阳550025

通信作者：王小利；E-mail: wangxiaolizhenyuan@126.com

摘　要：

以高羊茅(Festuca arundinacea) ‘黔草1号’为材料, 利用cDNA末端快速扩增(RACE)技术克隆了调控植物开花关键基因FLOWERING LOCUS T (FT), 命名为FaFT2。序列分析结果表明, FaFT2基因cDNA全长1 073 bp, 具有开放阅读框为537 bp, 编码一条含178个氨基酸残基的蛋白质。该蛋白序列比对发现, 所推测的FaFT2氨基酸序列具有1个保守的磷脂酰乙醇胺结合蛋白(PEBP)结构域。系统遗传进化树分析表明FaFT2与黑麦草、节节麦和二穗短柄草的FT亲缘关系较近。利用实时荧光定量PCR技术分析高羊茅苗期FaFT2表达水平, 发现该基因在光照时段表达水平持续上调, 黑暗时段表达水平下调。由此推测, FaFT2基因与高羊茅光周期敏感性具有一定的相关性, 可能在光周期调控开花途径中发挥作用。

关键词：高羊茅; FaFT2基因; 基因克隆; 生物信息学分析

收稿：2017-06-12   修定：2017-07-27

资助：国家自然科学基金项目(31360576)、贵州省农科院自主创新专项[字(2014)号]、贵州省科学技术基金(20132151)和贵州省科技厅重大专项[字(2014) 6017号]。

Cloning and expression analysis of FaFT2 gene in tall fescue

CHEN Xi^1,2, ZHAO De-Gang^1,2, CHEN Ying¹, LI Xiao-Dong¹,WU Jia-Hai¹, WANG Xiao-Li^1,2,*

¹Guizhou Institute of Prataculture, Guizhou Academy of Agricultural Sciences, Guiyang 550006, China; ²The National-Local Joint Engineering Research Center of Karst Region Plant Resources Utilization & Breeding of Guizhou Province, Guizhou University, Guiyang 550025, China

Corresponding author: WANG Xiao-Li; E-mail: wangxiaolizhenyuan@126.com

Abstract:

FLOWERING LOCUS T (FT) gene was cloned through rapid amplification of cDNA ends (RACE) technique from Festuca arundinacea cv. ‘Qiancao No.1’. The result of sequence analysis showed that the fulllength cDNA (1 073 bp) was obtained, named FaFT2, with a predicted 537 bp open reading frame, which encoded 178 amino acids. By BLSTP analysis, the conserved phosphatidyl ethanolamine-binding protein (PEBP) domain was found in FaFT2 protein sequence. Phylogenetic analysis showed that FaFT2 was closely related to FTs from other Gramineae plants such as Lolium perenne, Aegilops tauschii and Brachypodium distachyon. Quantitative real-time PCR assays showed the expression of FaFT2 in tall fescue seedling. The transcript levels of the FaFT2 gene was continuous up-regulated during the light period and down-regulated in the dark. The results suggest that FaFT2 is photoperiod sensitive and play a functional role in flowering regulation by photoperiod in tall fescue.

Key words: tall fescue; FaFT2 gene; gene cloning; bioinformatics analysis