玉米热激转录因子ZmHsf06的耐热功能及其调控通路研究  

Study on Thermotolerance Function and Regulatory Pathways of Heat Shock Transcription Factor ZmHsf06 in Maize(Zea mays)

作　　者：李冉 马贞玉[2] 张世昌 李国良[2] 孟祥照 段硕楠[2] 刘子会[2] 吕爱枝[1] 郭秀林[2] 张华宁[2] LI Ran;MA Zhen-Yu;ZHANG Shi-Chang;LI Guo-Liang;MENG Xiang-Zhao;DUAN Shuo-Nan;LIU Zi-Hui;LYU Ai-Zhi;GUO Xiu-Lin;ZHANG Hua-Ning(College of Agriculture and Forestry Science and Technology,Hebei North University,Zhangjiakou 075000,China;Institute of Biotechnology and Food Science/Hebei Key Laboratory of Plant Genetic Engineering,Hebei Academy of Agriculture and Forestry Sciences,Shijiazhuang 050051,China;Wheat Engineering Research Center of Hebei Province,Shijiazhuang Academy of Agriculture and Forestry Sciences,Shijiazhuang 050041,China)

机构地区：[1]河北北方学院农林科技学院,张家口075000 [2]河北省农林科学院生物技术与食品科学研究所/河北省植物转基因工程重点实验室,石家庄050051 [3]石家庄市农林科学研究院河北省小麦工程技术研究中心,石家庄050041

出　　处：《农业生物技术学报》2024年第3期505-517,共13页Journal of Agricultural Biotechnology

基　　金：河北省自然科学基金(C2021301038);河北省农林科学院基本科研业务费(2021110301)。

摘　　要：玉米(Zea mays)不同生育期遭遇高温热浪渐成常态,严重影响产量和品质。热激转录因子(heat shock transcription factor,Hsf)能够激活下游多种耐热途径,调控耐热相关基因表达从而提高植株耐热性。本研究在前期对玉米HsfA1亚族基因ZmHsf06特性及耐热性功能初步研究的基础上,通过创制ZmHsf06过表达玉米株系,深入研究该基因的耐热功能,进一步通过染色质免疫共沉淀测序(chromatin immunoprecipitation,ChIP-seq)和蛋白互作及转录激活活性实验,分析并确定ZmHsf06的下游靶基因和高富集通路。结果表明,ZmHsf06能自身互作形成同源多聚体,具有转录激活活性;与野生型相比,ZmHsf06过表达玉米株系的耐热性明显提高。ChIP-seq分析表明,热胁迫下共富集差异基因2218个,其中受ZmHsf06诱导特异上调的基因有1169个,主要富集在电子载体活性、光合作用、内质网蛋白加工和热激响应等通路中。ZmHsf06结合基序同时含有热激元件、C2C2、TEAD、MADS等其他转录因子的结合元件和一些未知元件。凝胶迁移实验表明,ZmHsf06蛋白与筛选出的3个代表性基因(Zm00001d000260,Zm00001d000272,Zm00001d033987)的启动子热激元件互作。本研究为深入研究玉米Hsf家族的耐热功能及其分子机制提供依据。Heat frequently affects yield and quality in different growth stages of maize(Zea mays).Heat shock transcription factor(Hsf)plays important roles in improving plant heat tolerance through regulating the expression of thermotolerance-related and activating multiple downstream pathways.Based on previous studies on characteristics and thermotolerance function of maize A1 subclass ZmHsf06,this study investigated its function and molecular mechanism in regulating thermotolerance through transformation in maize and chromatin immunoprecipitation-seq(ChIP-seq),and verified the protein interaction and transcriptional activation activity further.The results showed that ZmHsf06 had transcriptional activation activity and could interact with itself.Compared with the wild type,the thermotolerance of maize lines overexpressing ZmHsf06 were significantly improved.Analyzing the downstream target genes and high enrichment pathways of ZmHsf06 by ChIP-seq,a total of 2218 differentially enriched genes were found under heat stress,including 1169 genes up-regulated by ZmHsf06 specially,which were mainly enriched in electron carrier activity,photosynthesis,endoplasmic reticulum protein processing and heat shock response.The ZmHsf06 binding motif had heat shock elements(HSE),the binding elements of other transcription factors such as C2C2,TEAD and MADS and some unknown elements.Electrophoretic mobility shift assay(EMSA)showed that ZmHsf06 could bind to the heat shock elements located in the promoters of 3 target genes(Zm00001d000260,Zm00001d000272 and Zm00001d033987).These results will provide evidences for furthering the study of thermotolerance function and regulation mechanism of maize Hsf family.

关 键 词：玉米 热激转录因子 耐热性 染色质免疫共沉淀测序(ChIP-seq) 

分 类 号：S523[农业科学—作物学]