二穗短柄草BdGI基因表达及其蛋白互作分析

Analysis of BdGI Gene Expression and Protein Interaction in Brachypodium distachyon

光周期是调控植物开花的主要途径之一,GI在光周期途径中是控制生理节律和开花的关键因子。为探索二穗短柄草(Brachypodiam distachyon)BdGI基因的表达模式和蛋白互作关系,本研究通过实时荧光定量PCR(qRT-PCR)技术分析BdGI基因在不同光照条件下和长日照下不同生长发育阶段的表达情况;运用酵母双杂交初步筛选出互作蛋白BdZTL,并用双分子荧光互补法(BiFC)和免疫共沉淀法(CoImmunoprecipitation)进行验证。qRT-PCR分析结果表明,BdGI基因在不同光照下和长日照下不同生长发育阶段的表达情况不同,且都保持一定的昼夜节律,并且受到光周期的调控;利用酵母双杂交检测到GI与ZTL蛋白存在互作关系,双分子荧光互补与免疫共沉淀检测结果验证了两者互作关系的真实性。综上所述,BdGI的表达受光周期调控,具有昼夜节律性,在光周期诱导二穗短柄草开花的过程中也具备一定的调控作用。本研究结果为GI基因的功能研究奠定了理论基础。

Photoperiod is one of the main pathways regulating plant flowering, and GI is a key factor controlling circadian rhythm and flowering in the photoperiod pathway. To explore the expression pattern and protein interaction of the BdGI gene in Brachypodiam distachyon, quantitative real-time PCR(qRT-PCR) was used to analyze the expression of the BdGI gene under different light conditions and different growth stages at long sunshine. The interacting protein BdZTL was initially screened by yeast two-hybrid analysis and verified by BiFC and co-immunoprecipitation. qRT-PCR analysis showed that the expression of the BdGI gene was different in different growth and development stages under different light and long sunshine but maintained a certain circadian rhythm, and was regulated by photoperiod. The interaction between GI and ZTL proteins detected by yeast two-hybrid was vertified by fluorescence complementation and immunoprecipitation. In conclusion, these studies indicate that the expression of BdGI is regulated by photoperiod and has a circadian rhythm, it plays a certain regulatory role in the photoperiod-induced flowering of Brachypodium distachyon, which lays a foundation for further study on the function of this gene.