水稻miR156-SPL调控模块的生物信息学及高温胁迫下的表达  

Bioinformatics analysis and expression of miR156-SPL module in rice under heat stress

作　　者：骆鹰 陈蓉 黄珊珊 汪启明[2] 饶力群[2] 王伟平[3] 廖阳 LUO Ying;CHEN Rong;HUANG Shanshan;WANG Qiming;RAO Liqun;WANG Weiping;LIAO Yang(College of Chemistry and Biological Engineering,University of Science and Technology,Yongzhou 425199,China;College of Bioscience and Biotechnology,Hunan Agricult ural University,Changsha 410128,China;State Key Laboratory of Hybrid Rice,Hunan Hybrid Rice Research Center,Changsha 410125,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 [3]杂交水稻国家重点实验室,湖南杂交水稻研究中心,长沙410125

出　　处：《应用与环境生物学报》2024年第5期981-989,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：为解析miR156基因家族的进化特征、组织特异表达及其在水稻(Oryza sativa)应答高温胁迫下的调控作用,通过生物信息学对水稻osa-miR156家族进行鉴定,分析了miR156家族成员数量、染色体定位、序列特征、系统发育进化及其候选靶基因;基于psRNATarget数据库、结合降解组测序预测miR156l的靶基因,并利用qRT-PCR分析高温胁迫下osa-miR156l-5p及其靶基因的表达情况.结果表明,osa-miR156家族含有18条前体序列和25条成熟体序列,每条pre-MiR156前体序列均能形成稳定的茎环二级结构,其最小折叠自由能在-43.30-62.60 kal/mol范围;osa-miR156家族成员包括5个分支,与玉米(Zea mays)、大麦(Hordeum vulgare)的亲缘关系较近;水稻OsSPL家族成员共19个,在第2、6、8号染色体上的数量分布最多,均为3个,且OsSPL基因在水稻基因组中存在5对片段重复基因对;osa-MiR156前体基因启动子区域含生长发育、激素、应激响应等相关顺式作用元件,osa-miR156及其靶基因OsSPL家族成员在水稻不同组织中的表达均存在明显差异;靶基因预测显示,osa-miR156l的靶基因为OsSPL14、OsSPL16和OsSPL18,且osa-miR156l在与靶基因互补的第12个核苷酸处裂解靶标mRNA.qRT-PCR分析发现,高温胁迫下osa-miR156l-5p的表达水平呈先下降后上升再下降趋势,与靶基因OsSPL16表达呈负调控关系.本研究表明植物miR156家族序列特征具有高度保守性,水稻miR156-SPL途径可能在应答高温胁迫中起着重要的调控作用.(图9参34)The miR156 family is one of the most ancient and conserved families involved in the regulation of plant growth and development.However,the evolutionary characteristics,tissue-specific expression,and regulatory role of the miR156 gene family in the response of rice to high-temperature stress are unknown.In this study,bioinformatics was used to identify rice miR156 and its SPL gene family,and the number of miR156 family members,chromosome localization,sequence characteristics,phylogenetic evolution,and candidate target genes were analyzed.Target genes of miR156l-5p were predicted using the psRNATarget database combined with degradome sequencing.qRT-PCR was used to explore the expression levels of osa-miR156l and its target genes OsSPL14,OsSPL16,and OsSPL18 under heat stress.The results showed that the rice miR156 gene family contains 18 precursor sequences and 25 mature sequences,and each pre-MiR156 precursor sequence can form a stable stem-ring secondary structure with a minimum folding free energy ranging from -43.30 to 62.60 kal/mol.The phylogenetic tree of the osa-miR156 families includes five branches that are closely related to the miR156 of maize and barley.The OsSPL g ene f amily i n r ice c ontained 19 m embers,w ith t he l argest number(three)distributed on chromosomes 2,4,and 8.There were five pairs of repeated OsSPL fragments in the rice genome.The promoter regions of osa-MiR156 precursor genes contain cis-acting elements related to the photoresponse,transcription start sites,growth and development,hormones,and stress responses.The expression of osa-MiR156 and its target gene OsSPL varied widely in different r ice tissues.Based on t arget gene prediction analysis,the target genes of miR156l were SPL14,SPL16,and SPL18,and miR156l was found to cleave the target mRNA at the 12th nucleotide.qRT-PCR revealed that under high-temperature stress,the transcription level of rice osa-miR156l-5p first decreased,then increased,and then decreased again,with a negative regulatory relationship between osa-miR156l-5p an

关 键 词：水稻 miR156 进化特征 表达模式 SPL 

分 类 号：S511[农业科学—作物学]