基于Masc基因信号通路的罗氏沼虾性别调控及其分子机制研究

Macrobrachium rosenbergii (M.rosenbergii) is one of the most important aquatic economic prawns in China. At present, there are little known to the sex regulatory genes and the mechanism of sex regulation in prawns. The masculinizer　gene (Masc) family is an important component of upstream pathway of insect sex regulation cascade network. In this study, based on the study of Masc gene, the molecular cloning, structure analysis and spatiotemporal expression of Masc gene were improved by using the methods and techniques of molecular biology, such as fluorescence quantitative PCR, in situ hybridization and immunohistochemistry. Moreover, RNA interference and animal experiments were used to study the phenotypic changes in the process of sex induction, sex reversal, gonad remodeling and sexual character maintenance of M.rosenbergii after Masc gene silencing. At the same time, in order to explore the sex regulatory genes of molecular and regulation mechanism, the expression and modification changes of other related key genes and proteins in sex regulation pathway of M.rosenbergii were studied. Similar to multiple gene cascade regulatory pathways, such as Fem > Masc > Sxl > tra > Dmrt / dsx) ≧Mr-IAG﹥terminal differentiation genes, a Masc gene based signal pathway for sex regulation of M.rosenbergii and its cascade regulatory pathway were constructed, and the mechanism of sex regulation would be elucidated.

罗氏沼虾(Macrobrachiun rosenbergii) 是我国重要的水产经济养殖动物，其性别调控功能基因的发掘存在诸多空白。雄性化基因家族Masc是昆虫性别调控上游通路的重要组成。本项目以罗氏沼虾Masc基因的性别调控研究为切入点，利用荧光定量PCR、原位杂交和免疫组化等方法，开展Masc基因的分子克隆、结构解析和时空表达规律研究。然后，采用RNA干扰和动物实验相结合的方法，开展Masc基因沉默后其性别诱导、性别逆转、性腺重塑和性征维持过程中的表型变化研究。同时，结合基因沉默后，性别调控途径中关键性控基因和蛋白的表达、修饰变化情况，依照类似Fem> Masc>Sxl﹥tra﹥Dmrt (dsx)≧Mr-IAG﹥末端分化基因的多基因级联调控通路，完成1个基于Masc基因的罗氏沼虾性别调控信号途径及其级联调控通路的网络构架，基本阐明其在罗氏沼虾性别分化与性别维持中分子机制和调控机理。

罗氏沼虾(Macrobrachiun rosenbergii) 是我国重要的水产经济养殖动物，其性别调控功能基因的发掘存在诸多空白。本项目以罗氏沼虾性别调控通路的关键基因Mr-Masc为切入点，阐明罗氏沼虾Mr-Masc基因的结构解析和时空表达规律；从分子水平和动物实验两方面切入，阐明Mr-Masc基因沉默对性别分化、性别维持、性别诱导、性别逆转和性腺重塑的影响和作用，阐明其在罗氏沼虾中性别分化和性别维持中的作用机制；阐明基于罗氏沼虾Mr-Masc基因的性别决定和性别维持的网络调控通路，为虾类性别调控研究领域提供详实数据和新的研究思路；开展罗氏沼虾假雌个体的性腺发育和繁育特性研究，为Mr-Masc基因分子选育技术在全雄单性苗种中的应用提供实施例。项目实施过程中，发表虾类性别和生殖调控研究方向SCI论文6篇，申请虾类性别调控方向的发明专利1项，获授权实用新型专利1项，培养7名动物性别调控方向的研究生，保质超量完成研究目标。