20E早期应答基因及其在对虾生长中的调控机理研究

Molting, an important event in shrimp growth process, its researches have important theoretical value and economic significance. However, it is not complete about the molting signal regulation network. A main gap is how the 20-hydroxyecdysone (20E) response genes regulates downstream genes to control molting and growth in the ecdysone signal pathway. In our recent studies, many possible response genes and downstream genes in 20E signaling have found. In this project, we will use Litopenaeus. vannamei as the research materials, screen and identify the 20E early response genes (such as BR-C, E75, E74, E93, HR3, FTZ-F1 and so on) by RNA-seq and bioinformatics analysis. The functions of these genes will be analyzed through RNA interference and gene knockout technology, and their downstream target genes will be identified by ChiP-seq. Further, a downstream molting hormone signaling regulatory network will be established in shrimp and the mechanism of molting and growth will be explored. The results of this study will be able to illustrate the characteristics of shrimp growth and hormonal regulation, but also for the development and utilization of shrimp functional genes to improve shrimp aquaculture.

蜕皮是对虾生长过程中的一个重要事件，其研究具有重要的理论价值和经济意义。然而目前有关对虾蜕皮信号的调控网络尚不完整，突出的缺口是蜕皮激素信号传递至EcR/RXR后，蜕皮激素(20-hydroxyecdysone, 20E) 应答因子如何调控下游分子，进而表现出蜕皮和生长现象，其分子机理尚不明确。近期研究发现了多个可能的应答基因及20E信号通路下游分子。本课题拟以凡纳滨对虾为材料，通过RNA-Seq和生物信息学分析，筛选鉴定对虾20E早期应答基因如BR-C、E75、E74、E93、HR3、FTZ-F1等，通过RNA干扰和基因编辑技术分析其功能，通过ChiP测序等技术鉴定其下游靶基因，进而初步建立对虾蜕皮激素信号下游的分子调控网络；在此基础上重点探究对虾蜕皮和生长分子机理，挖掘对虾生长调控关键基因。本研究可为阐明对虾激素调控机理，开发利用功能基因、指导养殖和选育建立理论基础。

本课题在对虾基因组等多组学研究的基础上，筛选、鉴定和分析了凡纳滨对虾蜕皮激素20E受体和早期应答基因，包括EcR、RXR、E75、Br-C、HR3、Ftz-F1等，确定了各基因结构、表达特征以及系统发生特点；通过RNA干扰实验分析了20E受体和多个早期应答基因的功能：观察和分析了干扰后成体蜕皮生长情况以及相关基因表达情况；同时分析和鉴定了20E早期应答基因调控的下游多个重要的调控基因和效应基因，如GSK3、Sulf-1、IGFBP、Wnt、Myostatin、Actin、Myosin等，并分析了其结构和功能，初步构建了比较完整的蜕皮信号调控网络；在此基础上分析了对虾蜕皮信号通路与生长信号通路的关联，并探讨了这些基因在对虾的遗传育种中的可能的应用。本课题研究结果为深入了解虾类激素调节蜕皮和生长的机理，揭示对虾系统发生和以及分子育种提供新的线索和思路。