小麦TaGeBPL基因的克隆及表达分析  

Cloning and Expression Analysis of TaGeBPL gene in wheat

作　　者：王蒙蒙 张香宇 张露 徐晓敏 王琦 王长有[1] 吉万全[1,2] 张宏 WANG Mengmeng;ZHANG Xiangyu;ZHANG Lu;XU Xiaomin;WANG Qi;WANG Changyou;JI Wanquan;ZHANG Hong(College of Agronomy, Northwest A&F University, Yangling, Shaanxi 712100, China;Shaanxi Research Station of Crop Gene Resources & Germplasm Enhancement, Ministry of Agriculture, Yangling, Shaanxi 712100, China)

机构地区：[1]西北农林科技大学农学院,陕西杨陵712100 [2]农业部作物基因资源与种质创制陕西科学观测实验站,陕西杨凌712100

出　　处：《麦类作物学报》2021年第9期1063-1072,共10页Journal of Triticeae Crops

基　　金：陕西省科技创新团队项目(2018TD-004);陕西省重点研发计划项目(2019ZDLNY04-06);陕西省自然科学基金项目(2021JM-090)。

摘　　要：无表皮毛增强子结合蛋白GeBP(GLABROUS1 enhancer-binding protein)是植物体内特有的一类转录因子,可调控植物表皮毛的生长过程,在植物生长发育、衰老、抗逆、防御反应进程中发挥着重要作用。前期对自发斑点细胞坏死RILs(N13039H/N)进行转录组学比较,发现一个未知功能基因在两个材料中表达差异显著。为深入了解小麦中该基因的序列特征和表达特性,通过RT-PCR方法克隆获得了CDS区域片段。生物信息学分析结果表明,该基因位于3D染色体上,CDS编码区长度为1527 bp,共编码508个氨基酸,含有1个DUF573结构域和3个核定位信号。InterPro比对表明,编码蛋白属于GeBP家族,故将该基因命名为TaGeBPL。亚细胞定位结果表明,TaGeBPL定位于细胞核。白粉病菌(Blumeria graminis f.sp.tritici,Bgt)侵染条件下的定量qRT-PCR结果表明,TaGeBPL基因在感白粉病近等基因系N9134S中的表达模式呈双峰曲线变化,侵染12和36 h时,基因表达显著上调,而在抗白粉病近等基因系N9134R中的表达呈单峰曲线变化,在侵染12 h时显著下调。TaGeBPL在N9134S中的相对表达量始终高于在N9134R中的表达量,由此推测TaGeBPL基因可能参与病原菌响应的相关途径。酵母转录激活试验表明,TaGeBPL转录因子没有转录自激活活性。本研究结果为深入解析GeBP的功能奠定了基础。GeBP(GLABROUS1 enhancer-binding protein)is a plant specific transcription factor,playing important roles in plant growth,senescence,resistance and defense.Previous transcriptomics comparing RILs(N13039H/N),phenotyping spotted Vs.normal leaves,showed significant differences in the expression of an unknown functional gene in the two materials.In order to understand the sequence characteristics and expression patterns of this gene in wheat,the gene was cloned by RT-PCR method.The results showed that the length of CDS is 1527 bp,encoding 508 amino acids,which contains 1 DUF573 domain and 3 nuclear localization signals.InterPro indicated that the encoding protein belong to the GeBP family and the gene was designated as TaGeBPL.The subcellular localization results showed that TaGeBPL was localized in the nucleus.The results of qRT-PCR showed that the expression of TaGeBPL showed a double-peak curve in the powdery mildew-susceptible near-isogenic line N9134S and the expression of TaGeBPL were significantly up-regulated at 12 h and 36 h after inoculation in N9134S.While the expression of TaGeBPL showed a mono-peak curve in the powdery mildew resistant near-isogenic line N9134R.The expression level was significantly down-regulated at 12 h after inoculation in N9134R.And the relative expression level of TaGeBPL in N9134S was always higher than that in N9134R.It was speculated that TaGeBPL gene might be involved in the related pathway of pathogen response.The yeast transcription activation experiments indicated that the TaGeBPL transcription factor had no transcriptional self-activation activity.The results lay a foundation for further analysis of the GeBP function.

关 键 词：小麦 GeBP转录因子 亚细胞定位 表达分析 自激活 

分 类 号：S512.1[农业科学—作物学] S330