小麦TaDEP1基因启动子区转录增强序列的鉴定  

Identification of Transcription Enhancer in Promoter Region of TaDEP1

作　　者：谷玉娟 毛丰 路芳芳 刘斌 刘磊 高慧 GU Yujuan;MAO Feng;LU Fangfang;LIU Bin;LIU Lei;GAO Hui(Hebei Normal University of Science and Technology,Qinhuangdao,Hebei 066004;Jiangsu Key Laboratory for Eco Agricultural Biotechnology around Hongze Lake,Huaiyin Normal University,Huaian,Jiangsu 223300;Suzhou University of Science and Technology,School of Chemistry and Life Sciences,Suzhou,Jiangsu 215004)

机构地区：[1]河北科技师范学院,河北秦皇岛066004 [2]淮阴师范学院生命科学学院环洪泽湖生态农业生物技术重点实验室,江苏淮安223300 [3]苏州科技大学化学与生命科学学院,江苏苏州215004

出　　处：《核农学报》2022年第6期1100-1107,共8页Journal of Nuclear Agricultural Sciences

基　　金：国家自然科学基金资助项目(31700274);河北省自然基金资助项目(C2019407001);河北省高等学校科学技术研究项目(QN2020154);河北省在读研究生创新能力培养资助项目(CXZZSS2021152)。

摘　　要：RNAi干涉小麦TaDEP1基因导致小穗密度下降以及小穗数减少,表明该基因转录水平的降低会引起基因功能的丧失。为挖掘影响TaDEP1转录水平的作用元件,通过特异扩增D组TaDEP1基因启动子并进行测序,发现不同品种间存在132 bp序列差异,该序列位于TaDEP1基因起始密码子上游1529~1660 bp处。利用小麦微核心种质材料,鉴定到132 bp序列与TaDEP1基因的转录水平呈正相关,即启动子序列中含有132 bp序列的TaDEP1基因转录水平明显高于不含该序列的转录水平。启动子顺式作用元件分析结果显示,该132 bp序列未包含特殊的顺式作用元件。结合荧光素酶(LUC)活性试验,在小麦原生质体中证明该序列能够显著提高LUC的转录活性,表明该序列可作为小麦基因转录增强序列。本研究鉴定到新的小麦基因转录增强序列,为小麦分子标记辅助选择育种提供了基因资源与技术支持。Knock-down of TaDEP1 by RNA interference leads to a decrease in spikelet density and a decrease in spikelet number,indicating that decreasing the transcriptional level of TaDEP1 can result in the loss of its function.To uncover the cis-elements required for TaDEP1 transcriptional regulation,this study identified a genetic variation of a 132 bp sequences among different varieties via specifically amplifying and sequencing the TaDEP1 promoter in D subgenome,which is located at 1529~1660 bp upstream of the start codon of the TaDEP1 gene.Furthermore,it was observed that the 132 bp sequence is positively associated with the transcriptional level of TaDEP1 in wheat micro-core collection,that is,the transcriptional level of TaDEP1 gene with the 132 bp sequence within the promoter sequence was significantly higher than that without the 132 bp sequences.Bioinformatics analysis revealed that there was no annotated cis-elements in the 132 bp sequences.Based on the LUC activity assay,it was determined that this sequence can significantly increase the transcription activity of LUC in wheat protoplasts,indicating that this sequence can be used as a wheat gene transcription enhancer.This study identified a new wheat gene transcription enhancer sequence,and on the other hand,it also provided gene resources and technical support for wheat molecular marker-assisted selection breeding.

关 键 词：小麦 TaDEP1 启动子序列 转录水平 

分 类 号：S512.1[农业科学—作物学]