玉米花粉高温胁迫相关miRNA的筛选及其靶基因分析  

Screening of miRNAs Related to High Temperature Stress in Maize Pollen and Analysis of Their Target Genes

作　　者：李川[1] 张盼盼 张美微[1] 牛军 穆蔚林 郭涵潇 乔江方[1] LI Chuan;ZHANG Panpan;ZHANG Meiwei;NIU Jun;MU Weilin;GUO Hanxiao;QIAO Jiangfang(Cereal Crops Institute,Henan Academy of Agricultural Sciences,Zhengzhou 450002,China)

机构地区：[1]河南省农业科学院粮食作物研究所,河南郑州450002

出　　处：《河南农业科学》2024年第2期1-16,共16页Journal of Henan Agricultural Sciences

基　　金：河南省农业科学院自主创新基金项目(2023ZC006)。

摘　　要：以高耐热玉米品种郑单958、低耐热玉米品种先玉335为材料,以正常生长条件为对照,在花期进行高温胁迫,通过miRNA高通量测序筛选玉米花粉中的差异表达miRNA,然后预测其靶基因,并对靶基因的本体特征和代谢通路进行富集分析。结果表明,共筛选到818个miRNA前体序列。在郑单958高温胁迫花粉与对照花粉对比组(HT958 vs CK958)中共筛选到19个显著差异表达miRNA序列,其中15个miRNA序列上调表达,4个下调表达,3个miRNA序列达到极显著水平(P<0.01)。对这19个显著差异表达miRNA的靶基因进行预测,共获得了503个基因转录本,其富集较多的GO生物学过程条目分别为转录调控DNA-模板、微管生物学过程、磷酸化作用、RNA聚合酶Ⅱ正向调控转录过程、甲基化作用等,KEGG富集较显著的代谢通路分别是谷胱甘肽代谢、碳代谢、花生四烯酸代谢、糖酵解/糖异生、叶酸生物合成等。在先玉335高温胁迫花粉与对照花粉对比组(HT335 vs CK335)中共筛选到15个显著差异表达miRNA序列,其中7个miRNA序列上调表达,8个下调表达,1个miRNA序列达到了极显著水平(P<0.01)。对这15个显著差异表达miRNA的靶基因进行预测,共获得了454个基因转录本,其富集较多的GO生物学过程条目分别为转录调控DNA-模板、磷酸化作用、蛋白质磷酸化作用、蛋白质水解、DNA修复等,富集较显著的KEGG代谢通路分别是其他多糖降解、亚油酸代谢、代谢通路、硫胺素代谢、内质网内蛋白质加工过程等。在郑单958高温胁迫花粉与先玉335高温胁迫花粉对比组(HT985 vs HT335)中共筛选到85个显著差异表达miRNA序列,其中35个miRNA序列为上调表达,50个为下调表达,24个miRNA序列达到了极显著水平(P<0.01)。对这85个显著差异表达miRNA的靶基因进行预测,共获得了2 286个基因转录本,其富集较多的GO生物学过程条目分别为转录调控DNA-模板、磷酸化作用、蛋白质磷酸By using high‐throughput miRNA sequencing technology,differentially expressed miRNAs were indentified in pollens of high heat tolerant maize variety Zhengdan 958 and low heat tolerant maize variety Xianyu 335 under high temperature stress at flowering stage,their target genes were predicted,and the enrichment analysis of genetic characteristics and metabolic pathways of these target genes were conducted.The results showed that a total of 818 miRNA precursor sequences were identified.Nineteen significantly differentially expressed miRNA sequences were screened in the group of Zhengdan 958 pollen under high temperature stress compared with the control pollen under normal condition(HT958 vs CK958).Among them,15 miRNA sequences were up‐regulated,4 miRNA sequences were down‐regulated,and expression level difference of 3 miRNA sequences reached extremely significant level(P<0.01).The predicted results of target genes of 19 differentially expressed miRNA sequences showed that a total of 503 gene transcripts were obtained,their enriched GO biological process terms included regulation of transcription,DNA‐templated,microtubule‐based process,phosphorylation,positive regulation of transcription by RNA polymeraseⅡ,methylation and so on,and the most significantly enriched KEGG pathways were glutathione metabolism,carbon metabolism,arachidonic acid metabolism,glycolysis/gluconeogenesis,folate biosynthesis and so on.Fiveteen significantly differentially expressed miRNA sequences were identified in the group of Xianyu 335 pollen under high temperature stress compared with the control pollen(HT335 vs CK335).Among them,7 miRNA sequences were up‐regulated,8 were down‐regulated,and the expression level difference of 1 miRNA sequence reached highly significant level(P<0.01).The predicted results of target genes of 15 differentially expressed miRNA sequences showed that a total of 454 gene transcripts were obtained,their enriched GO biological process terms included regulation of transcription,DNA‐templated,phosphory

关 键 词：玉米 花粉 花期高温胁迫 miRNA测序 差异表达miRNA 靶基因 

分 类 号：S513[农业科学—作物学]