基于RNA-seq和PER-seq联合分析探究ZmHDZ6表达调控网络  

Integrative analysis of RNA-seq and PER-seq to elucidate regulatory network of ZmHDZ6 expression

作　　者：方应浩 周波[3] 陈茹梅[2] 杨文竹[2] 秦慧民[1] FANG Ying-Hao;ZHOU Bo;CHEN Ru-Mei;YANG Wen-Zhu;QIN Hui-Min(Key Laboratory of Industrial Fermentation Microbiology of the Ministry of Education/Tianjin Key Laboratory of Industrial Microbiology/College of Biotechnology,Tianjin University of Science and Technology/National Engineering Laboratory for Industrial Enzymes,Tianjin 300457,China;Crop Functional Genome Research Center,Biotechnology Research Institute,Chinese Academy of Agricultural Sciences,Beijing 100081,China;Cereal Crops Institute,Henan Academy of Agricultural Sciences,Zhengzhou 450002,Henan,China)

机构地区：[1]工业发酵微生物教育部重点实验室/天津市工业微生物重点实验室/天津科技大学生物技术学院/工业酶国家工程实验室,天津300457 [2]中国农业科学院生物技术研究所作物功能基因组研究中心,北京100081 [3]河南省农业科学院粮食作物研究所,河南郑州450002

出　　处：《作物学报》2025年第4期958-968,共11页Acta Agronomica Sinica

基　　金：国家自然科学基金项目(32372064)资助。

摘　　要：玉米是需水量较大的作物,而干旱是制约玉米生产的主要因素。结合前期研究基础和相关研究进展,发现ZmHDZ6受干旱诱导强烈,且过表达植株表现出优良的抗旱性能。为探究玉米转录因子ZmHDZ6的下游调控机制,通过对ZmHDZ6过表达转基因玉米株系进行RNA-seq测序,对自交系B73原生质体进行PER-seq(protoplast transient expression-based RNA-sequencing)测序,并进行联合分析。结果显示,2种测序策略得到的差异表达基因(DEGs)呈现一致性,功能主要集中在参与氧化还原反应等GO富集分析条目。KEGG分析显示功能都富集在苯并噁唑嗪酮类化合物代谢途径上。此外,基于RNA-seq的DEGs在氨基酸和核苷酸代谢途径富集,而基于PER-seq的DEGs还富集在核糖体生物发生代谢途径。因此,推测ZmHDZ6可能通过调控与氧化还原相关基因和苯并噁唑嗪酮类化合物的代谢进而增强玉米抗旱性。通过进一步在129个Co-DEGs(Common DEGs)的启动子序列扫描HDZIP I家族的DNA结合基序(motif),将潜在靶基因缩减至16个,其中8个基因的功能与玉米抗逆紧密相关。本研究利用转录组学数据分析了ZmHDZ6基因的下游表达调控网络,为进一步解析抗旱机制提供了参考。Maize is a crop with high water demand,and drought is a major factor limiting its productivity.Building on previous findings and related research,we identified that the ZmHDZ6 gene is strongly induced by drought,and transgenic plants overexpressing ZmHDZ6 exhibit enhanced drought resistance.To investigate the downstream regulatory mechanisms of the maize transcription factor ZmHDZ6,RNA-seq was performed on transgenic maize plants overexpressing ZmHDZ6,while PER-seq(protoplast transient expression-based RNA sequencing)was conducted using protoplasts from the B73 inbred line.An integrative analysis of these datasets revealed that the differentially expressed genes(DEGs)identified by both methods showed consistent results in GO analysis,with their functions primarily associated with redox reactions.KEGG pathway analysis further demonstrated consistency in the benzoxazinoid biosynthesis pathway.Notably,RNA-seq DEGs were additionally enriched in amino acid and nucleotide metabolism pathways,while PER-seq DEGs were enriched in ribosome biogenesis pathways.Based on these findings,we propose that ZmHDZ6 enhances drought resistance in maize by regulating genes involved in redox processes and benzoxazinoid metabolism.Furthermore,through an integrative analysis of DNA binding motifs of the HD-ZIP I family and the promoters of 129 common DEGs(Co-DEGs),combined with gene annotation and motif physical location information,we narrowed potential target genes down to 16,of which 8 are closely associated with stress responses in maize.This study provides a detailed analysis of the regulatory network of ZmHDZ6 expression,offering valuable insights into the drought resistance mechanisms mediated by ZmHDZ6 and a reference for further functional studies.

关 键 词：RNA-SEQ PER-seq 转录因子 ZmHDZ6 靶基因 MOTIF 

分 类 号：S51[农业科学—作物学]