水稻幼穗分化特异性启动子pRFL的克隆和特征分析  

Cloning and characterization of the rice RFL gene promoter with specific expression in young inflorescence tissue

作　　者：苏军[1] 林智敏[1] 刘霞[1] 胡太蛟[1] 李刚[1] 颜静宛[1] 

机构地区：[1]福建省农业科学院生物技术研究所福建省农业遗传工程重点实验室,福州350003

出　　处：《应用与环境生物学报》2016年第4期675-679,共5页Chinese Journal of Applied and Environmental Biology

基　　金：福建省科技计划项目(2015N0103);省属公益类科研院所科研专项(2014R1019-1)资助~~

摘　　要：组织特异性启动子能够驱动基因在特定的时期和部位表达,克服组成型启动子启动的外源基因在受体植物中非特异、持续、高效表达所造成的浪费,是基因工程技术最重要元件之一.本研究利用PCR技术从水稻基因组中克隆了幼穗分化特异表达基因RFL翻译起始位点上游2 001 bp的启动子序列,命名为pRFL.生物信息分析显示,该片段含有36个转录起始核心启动子元件TATA-box和多个启动子增强子区顺式作用元件CAAT-box,以及多个光反应调控元件和植物激素响应元件等.将其与GUS基因构建成植物表达载体,导入水稻"日本晴",组织化学染色法检测显示,转基因水稻植株叶片、茎均无GUS显色,花序及发育中的小花有较强表达;荧光定量PCR测定幼穗GUS基因转录活性,显示pRFL驱动的GUS基因表达量比actin启动子驱动的GUS基因表达量高2.9倍.上述结果初步证明了pRFL为幼穗分化特异性启动子.Tissue specific expression promoter is an essential element for genetic modified crops. It can drive gene expression exclusively in targeted tissues and in special period. RFL gene is recognized as a control in rice flowering time and plant architecture. In this study, we cloned and sequenced 2001 bp of the promoter region of the RFL gene, named pRFL. The pRFL region contains several cis-acting regulatory elements, including 36 core promoter elements around -30 of transcription start, 25 common cis-acting elements in promoter and enhancer regions, 11 cis-acting regulatory elements involved in light responsiveness, as well as multiple cis-acting elements involved in the plant regulator responsiveness. We ligated pRFL to the β-glucuronidase （GUS） reporter gene, and transformed it into a japonica rice cultivar to determine the expression pattern of pRFL. In transgenic rice lines carrying the GUS gene driven by the pRFL, GUS expression was detected in young inflorescence and developing spikelet, but not in leaves or stems. GUS expression level in young inflorescence was 2.9 times higher than that of transgenic rice carrying the GUS gene driven by the aetin promoter. These results suggest that pRFL is a promoter specific regulating gene expression in young inflorescence and developing spikelet.

关 键 词：水稻 幼穗分化 特异性启动子 克隆 特征分析 

分 类 号：Q943.2[生物学—植物学] S511[农业科学—作物学]