花鲈渗透调节关键基因的筛选及非编码RNA调控机制的研究

Salinity is one of the most important environmental factors that directly affect the survival, growth and physiological function of teleost fish. Osmoregulation is the most important process during adaptation to different salinity water. Understanding factors underlying the osmoregulation process has great agricultural implications in aquaculture fish species. Lateolabrax maculatus, which is euryhaline specie, enable comparative research in which one environment can be used as a standard in order to contrast the specific genes or pathways for saltwater or freshwater osmoregulation. In this proposal, we examined transcriptomes from osmoregulation-related tissues of L. maculatus under two different salinities to identify the genes and non-coding RNAs responsible for osmoregulation by using high-throughput deep sequencing technology. Our research plan to 1) analysis the gene expression pattern and identify differentially expressed genes, 2) identify lncRNAs and construct gene-lncRNA interaction network utilizing bioinformatics methods, 3) explore miRNAs, identify differential expressed miRNA and predict target genes, 4) taken together, to find the key genes, lncRNAs and miRNA which are responsible for osmoregulation and investigate the non-coding RNA-modulating networks underlying the osmoregulation. This study will provide theoretical basis for sustainable development of L. maculatus and other aquaculture species.

盐度是影响鱼类生存、生长发育及各项生理机能最重要的环境因子之一，而渗透调节是鱼类适应不同盐度环境最重要的调节过程。研究表明除了编码蛋白质的基因外，非编码RNA—microRNA（miRNA）及长链非编码RNA（lncRNA）对渗透调节基因具有重要的调控作用。因此本项目拟以广盐性的花鲈作为实验材料，利用生物信息学结合分子生物技术，比较海水和淡水中花鲈转录组中基因及非编码RNA的表达差异来解析其渗透调节的分子机制，以期达到以下目的：对转录组进行基因注释， miRNA测定及lncRNA鉴别；通过差异表达分析、共表达网络构建、靶基因预测及功能分析等获得渗透调节的核心调控模块、关键基因及关键非编码RNA；综合解析渗透调节过程中非编码RNA对基因的调控机制，阐明花鲈渗透调节的分子机制。本研究成果将有助于花鲈的苗种选育、规模化养殖及养殖区域扩大，为花鲈及其它水产鱼类养殖产业的可持续发展提供理论支撑。

盐度是鱼类生存环境中最重要的生态因子之一，与鱼类的繁殖发育、生存生长、体内各机能调节密切相关。鱼类应对盐度改变时，通过渗透调节机制来维持体内平衡，一系列参与渗透调节的功能基因及调控因子的表达方式发生改变。本研究主要以广盐性的花鲈为研究对象，对其渗透调节的分子机制进行研究。通过转录组测序技术，对不同盐度处理组的花鲈鳃、肝脏组织进行mRNA及miRNA的表达分析，筛选获得参与渗透调节的候选关键基因及关键miRNA；通过第三代PacBio单分子测序技术，获得了花鲈的全长转录组，并对可变剪接事件、lncRNA等进行鉴定；通过RNA-seq与Iso-seq测序数据联合分析，对可变剪接在花鲈渗透调控中的作用机制进行研究；此外，对渗透调节关键基因/基因家族进行全基因组鉴定及功能研究。本研究成果为全面解析花鲈的渗透调节机制奠定了基础，为鱼类的淡化、盐化养殖提供了理论依据。