水稻孕穗期高温胁迫相关miRNA的鉴定及其靶基因的预测  

Identification of miRNAs related to high-temperature stress in rice at the booting stage and prediction of their target genes

作　　者：骆鹰 廖阳 罗小芳 王韬[2] 汪启明[2] 饶力群[2] LUO Ying;LIAO Yang;LUO Xiaofang;WANG Tao;WANG Qiming;RAO Liqun(College of Chemistry and Biological Engineering,Hunan University of Science and Technology,Yongzhou 425199,China;College of Bioscience and Biotechnology,Hunan Agricultural University,Changsha 410128,China)

机构地区：[1]湖南科技学院化学与生物工程学院,永州425199 [2]湖南农业大学生物科学技术学院,长沙410128

出　　处：《应用与环境生物学报》2024年第4期802-810,共9页Chinese Journal of Applied and Environmental Biology

基　　金：湖南省自然科学基金项目(2018JJ2144,2023JJ50416);湖南省教育厅科学研究重点项目(18A480);永州市指导性科技计划项目(2022-YZKJZD-005)资助。

摘　　要：水稻(Oryza sativa)是世界上重要的经济和粮食作物之一,随着全球变暖及不断扩大范围,高温对植物所造成的损害是水稻种植区亟须解决的问题,miRNA及其靶基因在植物趋利避害、生长发育调节等扮演不可替代的角色.利用高通量测序技术构建水稻9311品种孕穗期高温胁迫0 h、3 h、6 h和12 h的小RNA文库并进行生物信息学分析.在水稻中共鉴定获取464个已知miRNAs和123个新miRNAs,差异表达miRNAs共25个,其中上调表达12个,下调表达13个,并预测到253个候选靶基因;GO分析发现,靶基因在生物过程、细胞组分、分子功能分别高度富集于转录调控、膜的组成、ATP结合条目等;KEGG分析表明,靶基因主要富集于代谢通路,为13个,与8个差异表达miRNAs相互作用,形成调控网络.通过荧光定量qRT-PCR技术对8个差异表达miRNAs及其靶基因进行表达验证,结果表明osa-miR164f、osamiR1850.1、osa-miR6249a和novel_miR_85对靶基因呈负调控关系,osa-miR2871a-5p和novel_miR_87对靶基因呈正调控关系.本研究表明水稻miRNA能够通过靶向基因表达应答高温胁迫,可为水稻高温胁迫调控机制研究提供相关miRNAs及其优良候选靶基因.(图8表1参31)Rice is an economically important food crop worldwide.The emergence and expansion of global warming present an urgent problem in rice-planting areas,as high temperatures can cause significant plant damage.Previous studies have shown that microRNAs(miRNAs)and their target genes play crucial roles in regulating plant growth and development.This study used high-throughput sequencing technology to construct small RNA libraries of rice 9311 at the booting stage after heat treatment for 0,3,6,and 12 hours.Bioinformatics analysis identified 464 known miRNAs and 123 novel miRNAs in rice,with 25 differentially expressed miRNAs—12 upregulated and 13 downregulated.Additionally,253 corresponding target genes were predicted.Gene Ontology(GO)analysis revealed that the target genes were highly enriched in transcription regulation,integral components of membranes,and ATP-binding items related to biological processes,cell components,and molecular functions.KEGG pathway analysis indicated that 13 target genes were enriched in metabolic pathways interacting with eight differentially expressed miRNAs to form a regulatory network.Further analysis using qRT-PCR of eight differentially expressed miRNAs and their target genes in response to high-temperature stress showed that osa-miR164f,osa-miR1850.1,osa-miR6249a,and novel_miR_85 had negative regulatory relationships with their target genes,whereas osa-miR2871a-5p and novel_miR_87 had positive regulatory relationships.Based on a comprehensive analysis,miRNAs in rice respond to high-temperature stress by targeting gene expression.This study identified relevant miRNAs and candidate genes crucial for understanding the regulatory mechanisms of high-temperature stress in rice.

关 键 词：水稻 高温胁迫 MIRNA 靶基因 表达调控 

分 类 号：S511[农业科学—作物学]