Molecular Cloning And Functional Study Of Hairy(Hry) From Nilaparvata Lugens(St?l)

Nilaparvata lugens is an important migratory pest in rice production,which seriously affects the yield and quality of rice in China and Southeast Asian countries.Juvenile hormone(JH)and Ecdysone(Ec)play an important role in the growth and development of Nilaparvata lugensr.The two hormones regulate the expression of downstream genes through their respective receptor proteins and corresponding signal transduction pathways in vivo,thereby affecting the growth and development of Nilaparvata lugens.Hairy(Hry)is a downstream gene regulated by juvenile hormone and its receptor Methoprene-tolerant(Met).In this paper,Hairy was cloned from Nilaparvata lugens and analyzed by bioinformatics.It was found that there were conservative basic Helix-Loop-Helix(bHLH)domain,Orange and WRPW region in the predicted protein sequence.Quantitative fluorescent PCR was used to study the temporal and spatial expression characteristics of Hairy in Nilaparvata lugens.It was found that Hairy had significant temporal and spatial expression characteristics.For example,Hairy had relatively high expression levels in eggs,wings and feet.Through RNA interference experiments,we found that Hairy is closely related to hormone signaling pathway,and has a significant impact on survival,molting,genital and wing development and ovarian development.Down-regulation of Hairy gene reduces the survival rate to a certain extent,increases the mortality rate during molting,and results in abnormal development of genitals and wings,as well as ovarian malformation.Co-Immunoprecipitation assay showed that Hairy protein interacted with r ppel-homologue 1(r-h1)and E93,the key transcription factors downstream of two hormone signaling pathways,and Met,the juvenile hormone receptor,respectively.Then the co-localization of Hry with r-h1,E93 and Met was found by immunofluorescence.Finally,transcriptome sequencing and quantitative PCR validation were performed on the samples after RNAi.Hairy was found to regulate the expression of related genes in glycolysis pathway and epidermal protein genes.This study not only helps us understand the signaling pathways of juvenile hormone and ecdysone,but also provides a theoretical basis for the discovery of novel biopesticide targets.