高羊茅FaFT基因表达、蛋白互作及生物学功能分析  

Expression,Protein Interactions and Biological Function Analysis of FaFT in Festuca arundinacea

作　　者：罗维 牟琼 舒健虹[2] 吴佳海 王小利[2] LUO Wei;MU Qiong;SHU Jian-hong;WU Jia-hai;WANG Xiao-li(Key Laboratory of Genetics,Breeding and Reproduction of Plateau Mountain Animals,Ministry of Education,College of Animal Science,Guizhou University,Guiyang 550025;Institute of Prataculture,Guizhou Academy of Agricultural Sciences,Guiyang 550006;Institute of Fruit Research,Guizhou Academy of Agricultural Sciences,Guiyang 550006)

机构地区：[1]贵州大学动物科学学院高原山地动物遗传育种与繁殖教育部重点实验室,贵阳550025 [2]贵州省农业科学院草业研究所,贵阳550006 [3]贵州省农业科学院果树科学研究所,贵阳550006

出　　处：《生物技术通报》2021年第4期8-17,共10页Biotechnology Bulletin

基　　金：国家自然科学基金项目(31860674);贵州省高层次创新型人才培养项目[黔科合平台人才(2018)5634,黔科合人才(2016)4024];贵州省科技计划项目[黔科合平台人才(2020)5005]。

摘　　要：FLOWERING LOCUS T(FT)对植物开花有重要的调控作用。为探索高羊茅FaFT基因的生物学功能,从前期高羊茅转录组中克隆得到FaFT基因。qRT-PCR分析表明FaFT基因在长日照、短日照、连续光照、连续黑暗、昼夜颠倒和长日照下不同发育阶段都保持一定的昼夜节律并受光周期的调控。亚细胞定位显示FaFT基因编码的蛋白位于细胞核中;利用酵母双杂交筛选出可能与FaFT互作的FD蛋白,并进一步通过双分子荧光互补(BiFC)方法进行验证FaFT与FD蛋白互作的真实性。通过构建过表达载体和功能补偿发现在长日照条件下FaFT基因促进拟南芥提前开花,FT基因功能补偿后恢复拟南芥突变体开花表型,突变体植株本身没有开花现象。以上结果说明高羊茅FaFT基因的生物学功能得以成功验证。FLOWERING LOCUS T(FT)plays a significant role in regulating plant flowering.In this study,the FaFT gene was cloned from the previous tall fescue’s transcriptome in order to explore the FaFT gene biological function.qRT-PCR analysis showed that the FaFT gene maintained a certain circadian rhythm and was regulated by the light cycle in different developmental stages under long-day,shortday,continuous light,continuous darkness,day-night inversion,and long-day light.Subcellular localization revealed that the FaFT protein was located in the nucleus.FD(FLOWERING LOCUS D)protein that may interact with FaFT was selected by yeast two-hybrid,and the authenticity of the interaction between FaFT and FD protein was further verified by the bimolecular fluorescence complementation(BiFC)method.Through the construction of overexpression vectors and functional compensation,it was found that FaFT gene promoted Arabidopsis early flowering under long-day condition.After functional compensation of FT gene,the flowering phenotype of Arabidopsis mutants was restored,and the mutant plants themselves showed no flowering phenomenon.The above results indicate that the biological function of the FaFT gene of F.arundinacea has been successfully verified.

关 键 词：高羊茅 FT基因 蛋白互作 生物功能 

分 类 号：S688.4[农业科学—观赏园艺]