摘要
为探究高羊茅FaGI基因的生物学功能,本研究利用酵母双杂交、双分子荧光互补和免疫共沉淀探究与FaGI互作的蛋白;通过农杆菌介导法将过表达载体p1300-FaGI遗传转化拟南芥,获得转FaGI基因拟南芥株系,以拟南芥野生型Col-0、过表达FaGI基因株系和gi突变体为材料进行转录组学测序并观察其开花表型。结果表明,利用酵母双杂交方法筛选出与FaGI互作的FaCO蛋白,并通过双分子荧光互补和免疫共沉淀证明了FaGI和FaCO在体内和体外存在互作关系;过表达FaGI基因拟南芥植株的开花时间比野生型Col-0提前约1.24 d;将FaGI-OE、gi与野生型比对,分别筛选出1963和92个差异表达基因(DEGs),与野生型植株相比,过表达FaGI基因株系的差异基因富集在与生长发育、光周期途径、激素合成和信号传导、碳代谢等相关生物过程和代谢通路。综上,FaGI影响光周期途径相关基因的表达,在长日照条件下过表达FaGI基因促进了拟南芥开花,同时该基因的功能具有多样性与复杂性,可作为高羊茅调控分子育种的目标基因。本研究结果为揭示FaGI基因的功能及其调控网络奠定了基础。
In order to explore the biological function of the FaGI gene in tall fescue.In this study,yeast two-hybrid,bimolecular fluorescence complementation and co-immunoprecipitation were used to explore the proteins interacting with FaGI,and the overexpression vector p1300-FaGI was transformed into Arabidopsis thaliana by an Agrobacterium-mediated method,and the FaGI transgenic Arabidopsis lines were obtained.The wild type Col-0,overexpression FaGI gene lines and gi mutant of Arabidopsis thaliana were used for transcriptome sequencing and observing flowering performance.The results showed that the FaCO protein interacting with FaGI was screened by yeast two-hybrid method,and the interaction between FaGI and FaCO in vivo and in vitro was proved by bimolecular fluorescence complementation and co-immunoprecipitation.The flowering time of Arabidopsis plants overexpressing the FaGI gene was about 1.24 days earlier than that of wild-type Col-0.Differentially expressed genes(DEGs)of 1963 and 92 were screened by comparing FaGIOE and gi with WT,respectively.Compared with wild-type plants,the differentially expressed genes of overexpression FaGI lines were enriched in biological processes and metabolic pathways related to growth and development,photoperiod pathway,hormone synthesis and signal transduction,and carbon metabolism.In conclusion,FaGI affects the expression of photoperiod pathway-related genes,and overexpression of FaGI promotes flowering in Arabidopsis under long-day light conditions.At the same time,the function of FaGI are diverse and complex,which can be used as target genes for molecular breeding of tall fescue.The results of this study laid the foundation for revealing the function of FaGI gene and its regulatory network.
作者
路雪萍
马培杰
童伟杨
罗文举
李亚娇
赵德刚
王小利
LU Xueping;MA Peijie;TONG Weiyang;LUO Wenju;LI Yajiao;ZHAO Degang;WANG Xiaoli(Key laboratory of Plant Resource Conservation and Germplasm Innovation in Mountainous Region(Ministry of Education),College of Life Sciences/Institute of Agro-bioengineering,Guizhou University,Guiyang,Guizhou 550025;Guizhou Institute of Prataculture,Guizhou Academy of Agricultural Sciences,Guiyang,Guizhou 550006;Department of Grassland Science,College of Animal Sciences,Guizhou University,Guiyang,Guizhou 550025)
出处
《核农学报》
CAS
CSCD
北大核心
2023年第1期27-41,共15页
Journal of Nuclear Agricultural Sciences
基金
国家自然科学基金(32060394)
贵州省高层次创新型人才培养项目[黔科合平台人才(2018)5634-2]
贵州省高层次创新型人才培养项目[黔科合人才(2016)4003]
贵州省科技计划项目[黔科合平台人才(2020)5005]。
关键词
高羊茅
FaGI基因
蛋白互作
异源表达
转录组学
Festuca arundinacea
FaGI gene
protein interaction
heterologous expression
transcriptomics
