水稻AP2/ERF转录因子基因OsERF096启动子克隆及活性分析  

Cloning and activity analysis of promoter of an AP2/ERF transcription factor gene OsERF096 in rice

作　　者：陈悦 陈茜 董伟峰 才晓溪 沈阳 杨珺凯 贾博为 孙明哲 孙晓丽 CHEN Yue;CHEN Xi;DONG Weifeng;CAI Xiaoxi;SHEN Yang;YANG Junkai;JIA Bowei;SUN Mingzhe;SUN Xiaoli(Crop Stress Molecular Biology Laboratory,College of Agriculture,Heilongjiang Bayi Agricultural University,Daqing 163319,China)

机构地区：[1]黑龙江八一农垦大学农学院作物逆境分子生物学实验室,黑龙江大庆163319

出　　处：《土壤与作物》2022年第2期159-169,共11页Soils and Crops

基　　金：国家自然科学基金青年项目(32101672);中央支持地方高校改革发展资金人才培养支持计划项目,黑龙江八一农垦大学研究生创新科研项目(YJSCX2021-Y41);中国博士后基金资助项目(2021T140185,2020M670929);黑龙江省博士后基金资助项目(LBH-Z20028).

摘　　要：提高水稻耐冷性对保障水稻高产、稳产具有重要意义。AP2/ERF转录因子能够抵抗外界逆境胁迫,课题组前期研究发现水稻OsERF096基因显著受冷胁迫诱导表达。本研究克隆了OsERF096启动子序列(-1~-1215 bp),通过软件预测发现OsERF096启动子中含有多个与低温及激素响应相关的顺式作用元件。根据元件分布特征进行了5′端片段缺失设计,构建了GUS表达载体P_(OsERF096-1215)-GUS、P_(OsERF096-1102)-GUS、P_(OsERF096-827)-GUS、P_(OsERF096-501)-GUS,并转化拟南芥。通过GUS染色和GUS基因qRT-PCR检测发现,正常生长条件下,4个不同长度的启动子中P_(OsERF096-1215)启动子活性最高,P_(OsERF096-1102)最低。与前期qRT-PCR检测不同,冷处理下OsERF096启动子活性未发生显著变化。这些结果为后续OsERF096基因的表达调控及生物学功能研究奠定重要基础。Enhancing chilling resistance is of significance to high and stable yield in rice.AP2/ERF transcription factor can resist the cold stress.Our previous study found that rice OsERF096 gene expression was significantly induced by cold stress.In this study,we cloned the OsERF096 promoter(-1~-1215 bp)and found several cis-elements related to cold and hormone responses.Based on the distribution of cis-elements in OsERF096 promoter,four 5′-truncated fragments were designed and cloned into the pCAMBIA3301 vector,resulting in P_(OsERF096-1215)-GUS,P_(OsERF096-1102)-GUS,P_(OsERF096-827)-GUS,P_(OsERF096-501)-GUS constructs,which were then transformed into Arabidopsis.By comparing the GUS staining and GUS gene expression analyzed by qRT-PCR,we found that under control conditions,among the four OsERF096 promoter fragments,P_(OsERF096-1215) and P_(OsERF096-1102) displayed the highest and lowest activity,respectively.Unexpectedly,no significant difference was observed in the activity of four OsERF096 promoter fragments under cold stress,which was contradictory to previous qRT-PCR result showing that OsERF096 expression was greatly induced by cold stress.The findings will facilitate further research on the regulatory mechanism of OsERF096 expression and biological function.

关 键 词：水稻 AP2/ERF OsERF096 启动子 GUS活性 

分 类 号：S511[农业科学—作物学]