(USP) Gene And Ecdysone Receptor (EcR) Gene On The Growth And Development Of Quercus

Steroid hormones regulate many developmental processes in higher organisms, including postembryonic development, metamorphosis, and reproduction.In the process of insect metamorphosis, ecdysteroid signal was transducted and amplificated by hormone through a series of transcription factors, thus completing the regulation of the growth and metamorphosis. Steroid hormone20-Hydroxyecdysone (20E) as the main active forms of insect ecdysteroid, starting the insect molting,20E has the function of transcriptional regulation when binds to two members of the nuclear receptor superfamily, namely the ecdysone receptor (EcR) and ultraspiracle protein (USP).20E can interact with the formation of heterologous, starting transcription factor expression and resulting in insect molting. Ecdysone receptor as the only one nuclear receptors possessing insect ligands, which is highly conserved. Ecdysteroid exerts its biological effects through specific ecdysone receptor, which widely expressed in the whole development of the insect.Ultraspiracle protein as a ligand activated transcription factor, through hydrophobic molecules with the small binding to regulate gene expression, which belongs to the nuclear receptor superfamily. It has the the typical structure which similar with the Ecdysone receptor structure, including the A/B domain, C domain, D domain and E domain, but without highly conserved F domain.USP as a candidate receptor of Juvenile Hormone, however USP ligands have not be identified so far.This study is divided into two parts, the Ultraspiracle protein (USP) gene function research of Polyrhachis vicina Roger and the ecdysone receptor (EcR) gene function research of Teleogryllus emma Ohmachi&Matsumura. l.The Ultraspiracle protein (USP) gene function research of Polyrhachis vicina Roger In this study, Polyrhachis vicina is used as the research material, Polyrhachis vicina is a social insect with gregarious life typical of polymorphism, and extensive and highly complex nervous system. It is a good experimental material for the studies of insect growth and development.In order to explore the distribution of Ultraspiracle gene USP in Polyrhachis vicina’s brain and the effction about the regulation of development. RNAi and fluorescence in situ hybridization were used to discuss the the correlation between USP and EcR gene in the adult.The results are offered as follows:1.1The analysis of PvUSP gene localization and quantitative The relevant research of Polyrhachis vicina USP gene in the brain of the distribution and quantitative by using Fluorescence in situ hybridization and real-time fluorescence quantitative PCR technique.Fluorescence in situ hybridization showed USP was widely expressed in the brains of Polyrhachis vicina from different castes, especially in mushroom body which may related to the Polyrhachis vicina duty and their nervous system functions of brain. The workers olfactory fiber cell relative to female (male) ant at high expression which may be related to the social division of labor, PvUS? gene may be involved in the construction workers olfactory information processing capacity; in females mushroom body Kenyon cells, the central complex, lateral protocerebrum was positive, which may be related to the female ant in the main exercise mating, spawning, reproduce social duties, PvUSP gene may be involved in the construction of the visual, learning and memory and courtship behavior in males; optic lobe relative high expression which may be related to male visual development, PvUSP may be involved in male visual nervous system construction. And the same detection by real-time fluorescent quantitative results also show that, the expression of PvUSP in different grades are different, the highest in male workers, the female, at least,associated with the diversity of Polyrhachis vicina caste differentiation and activities.It suggested that PvUSP related to the Polyrhachis vicina castes duty and their nervous system functions.1.2The gene expression of PvUSP and PvEcR Through the RNAi method, Polyrhachis vicina USP gene was silenced by vitro synthesis of PvUSP dsRNA to explore its effect on the growth of Polyrhachis vicina.Amplified fragment size is621bp, the double control experiment are blank and negative controls (GFP dsRNA, amplified fragment is328bp),10ng/μLconcentration with different grade to Polyrhachis vicina, at the end of feeding period the different grades of Polyrhachis vicina adult USP and EcR mRN A expression were determined, the results showed that the expression of Polyrhachis vicina USP gene in three grade were obviously decreased, compared with the control group exhibited significant difference (P<0.01), RNA interference experiment obviously, while the relative expression of PvUSP expression decreased, PvEcR expression levels all increased in different castes,especially for female body. The expression of female is distinct (P<0.05).Those indicate that PvEcR may be inhibited by PvUSP.2.The ecdysone receptor (EcR) gene function research of Teleogryllus emma Ohmachi&Matsumura. In this study, taking Teleogryllus emma four instar larvae as experiment material, using the RNAi technology, in order to explore the ecdysone receptor gene on the growth and the correlation with estrogen related receptor (ERR), the results are offered as follows: Using RNAi method, the Teleogryllus emma four instar larvae of EcR gene was silenced, to explore its effect on the growth and development of Teleogryllus emma. In vitro synthesis of TeEcR dsRNA, amplified fragment555bp, set the negative control (GFP dsRNA, amplified fragment is328bp),5ng/μLconcentration injection, the results show that with the passage of time TeEcR mRNA expression level decreased continuously, but TeERR mRNA expression level has no significant change. And the female wing end plate dysplasia, incomplete, that TeEcR is silent, ecdysone signaling pathway is blocked, molting TeEcR on Teleogryllus Emma metamorphosis,a regulatory role in wing developmentAnd assume that TeEcR may be in the upstream of TeERR,the correlation between TeEcR and TeERR is not remarkable in function of regulation.