BdCDF1基因表达模式分析和亚细胞定位分析

Analysis of BdCDF1 Gene Expression Pattern and Subcellular Localization

光周期是植物开花调控中的主要途径之一,CDF1则是光周期调节植物开花的关键基因。为了研究二穗短柄草BdCDF1基因在不同光照下的表达模式和CDF1的亚细胞定位,本研究采用二穗短柄草为材料,利用荧光定量PCR技术研究BdCDF1在植物不同发育时期和不同光照条件下的表达情况;通过GFP融合蛋白表达法观察BdCDF1蛋白的亚细胞定位。荧光定量PCR结果表明,BdCDF1在不同光照长度下和不同的生长发育时期下表达情况不同,在长日照处理和短日照处理下,BdCDF1的表达高峰都出现在ZT24这个时间点上,表现出了一定的昼夜节律;在转入连续光照后,BdCDF1的表达高峰又都转移至ZT20这个时间点,且黑暗处理下BdCDF1的表达水平整体上要小于光照处理下的表达水平,这体现了BdCDF1的表达在一定程度上受到光周期的影响;亚细胞定位结果显示BdCDF1蛋白位于细胞核中。本研究结果表明BdCDF1基因表达受到昼夜节律和光周期的共同调控,暗示BdCDF1在二穗短柄草的光周期途径调控开花的过程中发挥重要作用,为后续BdCDF1基因的功能验证打下基础。

Photoperiod is one of the main pathways in plant flowering regulation,and CDF1 is a key gene in photoperiod regulation of plant flowering.In order to study the expression pattern of BdCDF1 gene in Brachypodium distachyon under different light and subcellular localization.The expression of BdCDF1 at different developmental stages and under different light conditions was studied by fluorescence quantitative PCR.The subcellular localization of BdCDF1 protein was observed by GFP fusion protein expression method.Fluorescence quantitative PCR results showed that the expression of BdCDF1 was different under different light lengths and different growth and development periods.The peak of BdCDF1 expression appeared at ZT24 under both long and short day treatments,showing a certain circadian rhythm.After continuous illumination,the expression peaks of BdCDF1shifted to ZT20,and the expression level of BdCDF1 under dark treatment was lower than that under light treatment on the whole,indicating that the expression of BdCDF1 was affected by photoperiod to some extent.Subcellular localization showed that BdCDF1 protein was located in the nucleus.The results of this study showed that the expression of BdCDF1 gene is regulated by circadian rhythm and photoperiod,suggesting that BdCDF1plays an important role in the photoperiod regulation of floweringin Brachypodium distachyon,laying a foundation for the subsequent functional verification of BdCDF1 gene.