油茶sRNA分析及其对光合油脂代谢的调控  

sRNA analysis and regulation of photosynthesis and lipid metabolism of Camellia oleifera

作　　者：王保明 尹迎峰 颜廷骥 张元华 陈永忠[4] WANG Baoming;YIN Yingfeng;YAN Tingji;ZHANG Yuanhua;CHEN Yongzhong(College of Agriculture&Forestry Science and Technology,Hunan Applied Technology University,Changde 415000,Hunan,China;Department of Modern Agriculture,Linyi Vocational University of Science and Technology,Linyi 276000,Shandong,China;Linyi Talent Working Group,Linyi 276000,Shandong,China;National Engineering Technology Research Center of Oil-tea Camellia,Hunan Academy of Forestry,Changsha 410004,Hunan,China)

机构地区：[1]湖南应用技术学院农林科技学院,湖南常德415000 [2]临沂科技职业学院现代农业系,山东临沂276000 [3]临沂人才工作集团,山东临沂276000 [4]湖南省林业科学院国家油茶工程技术研究中心,湖南长沙410004

出　　处：《经济林研究》2024年第3期25-35,119,共12页Non-wood Forest Research

基　　金：湖南省教育厅科学研究重点项目(19A360);湖南省“十四五”应用特色学科(林学)(湘教通[2022]351号)。

摘　　要：【目的】分析鉴定油茶s RNA的种类、数量和特征,揭示mi RNA及靶基因的功能,为利用mi RNA及靶基因进行油茶遗传改良提供理论依据。【方法】以优良油茶品种‘横冲89’为材料,采集嫩叶和成熟种子,提取总RNA,磁珠分离富集m RNA,利用Illumina RNA-seq高通量测序技术构建s RNA文库,分析s RNA的结构特征、种类和数量、公共序列及特有序列。将s RNA定位到油茶转录组上,分析比对上的总s RNA种类和数量,进行已知mi RNA比对分析和首位碱基偏好性分析。利用mi REvo和mirdeep2软件分析新mi RNA的前体和成熟体,预测新mi RNA,进行mi RNA家族分析;进行mi RNA表达与差异分析、层次聚类分析。最后,进行mi RNA靶基因预测、候选靶基因的GO功能富集分析和KEGG通路富集分析以及参与光合作用、油酯代谢mi RNAs的表达分析和靶基因筛选。【结果】分别构建了油茶叶和种子的s RNA文库,18~40个核苷酸(nucleotide,nt)的净读长分别为14256189条和10822313条,属于6138179和4980518种s RNA。其中,24-nt sRNA最多。叶和种子s RNA的公共序列为773647种,特有序列分别为4206871种和5364532种。在两个s RNA文库中,比对上油茶转录组参照序列的总读长分别为5855009和4122070条,比对上的mi RNA前体为118种,mi RNA成熟体为78个。研究发现:18~23 nt已知mi RNA首位碱基多偏好U。鉴定出54个新mi RNA,22个已知mi RNA家族,包括105个已知家族成员;67个表达差异mi RNA,调控1346个靶基因。其中,26个mi RNAs为显著差异表达,调控644个靶基因。聚类分析表明:显著差异表达的mi RNA被分为2族,一族在叶和种子中表达较低,另一族则表达较高。GO分析揭示差异表达mi RNA的候选靶基因在生物学过程、细胞组分、分子功能3大类的51个条目中富集。KEGG分析表明:差异表达mi RNA的候选靶基因在萜类主链生物合成、甘油酯代谢、氨酰t RNA生物合成等代谢过程中显著富集。此外,研究�【Objective】The types,quantities and characteristics of sRNAs in Camellia oleifera were analyzed and identified,the miRNA’s functions and corresponding target genes were revealed,and the theoretical basis for genetic improvement of C.oleifera was provided by miRNAs and their target genes.【Method】The excellent C.oleifera cultivar‘Hengchong 89’were used as materials.Its tender leaves and mature seeds were collected to extract total RNA,and separated to enrich mRNA with magnetic beads.The Illumina RNA-seq high-throughput sequencing technology was used to obtain sRNA libraries.The structure characteristics,types,quantities,common and unique sequences of sRNAs were analyzed.Then,sRNAs were located to C.oleifera transcriptome to analyze types and quantities of mapped sRNAs.Mapped analysis and the first base bias analysis of known miRNAs were also carried out.Novel miRNAs were predicted by miREvo and miRDeep2 software.The miRNA family analysis,miRNA expression and differential analysis,clustering analysis were implemented.Finally,prediction of miRNA target genes,GO functional enrichment and KEGG pathway enrichment analysis of target gene candidates were also implemented.The analysis of differentially expressed miRNAs and screening target genes participated in photosynthesis and lipid metabolism were carried out.【Result】The leaf and seed sRNA libraries of C.oleifera were constructed.The clean read counts of 18-40 nt sRNAs were 14256189 and 10822313,respectively,belonging to types of 6138179 and 4980518,respectively.Among them,counts of 24-nt sRNA was the largest.The common sequence types of in the two sRNA libraries were 773647,while the unique sequences were 4206871 and 5364532,respectively.The total reads aligned to the reference sequences of C.oleifera transcriptome were 5855009 and 4122070,respectively in the two sRNA libraries.118 miRNA precursors and 78 matures were mapped.It was found that the first nuclotied bias of 18-23 nt known miRNAs was U.54 new miRNAs were identified,22 known miRNAs familie

关 键 词：油茶 SRNA MIRNA 靶基因 调控 光合油脂代谢 

分 类 号：S601[农业科学—园艺学] S794.4