Effects Of Juvenile Hormone Binding Protein On Growth And Development In Mythimna Separata

Juvenile hormone binding protein(JHBP)is a type of carrier proteins,it binds juvenile hormone(JH)to form a complex and transports juvenile hormone to its target organs.Then regulate the growth and development of insects.In order to explore the functional role of JHBP in Mythimna separata,we cloned 7 Ms JHBP c DNA sequences,measured sequence expression by real-time PCR technology,interfered Ms JHBP3 genes by RNA interference technology,and explored the effect of interference JHBP c DNA sequence on the transcriptome of M.separata through transcriptome data analysis.The main findings are as follows:(1)Seven juvenile hormone binding proteins c DNA sequences were obtained from the M.separata,all sequences complete open reading frame(ORF).7 sequences were named Ms JHBP1-7,and they were logged on NCBI Gen Bank with the entry number MZ577066-MZ57072.The length of the 7sequences were 1190,731,839,816,1038,980 and 630 bp,respectively.Each sequence contained an open reading frame with length of 759,699,729,777,783,786 and 615 bp,encoding 252,232,242,258,260,261 and 204 amino acids,respectively.All of these c DNA sequences belong to the juvenile hormone binding protein family,and all contain the protein structure of the typical b HLH-PAS transcription factor family.(2)Juvenile hormone analogues(JHA)have different induction effects on different Ms JHBP c DNA sequence.Ms JHBP5 was significantly up-regulated after 48 h and Ms JHBP7 was significantly up-regulated after 12 h.After JHA treatment,Ms JHBP3 was up-regulated up to 38.55 fold and Ms JHBP5 was 20.60 fold,the induction effect of the other 5 genes was not significant,so these 2sequences were selected for subsequent experiments.At low concentrations,3 of the 7 sequences were significantly induced.When the concentration increases,the induction effected decreases,or there was no induction effect,even the inhibition effect appears.(3)The expression of Ms JHBP3 and Ms JHBP5 in different developmental stages of M.separata was different.Ms JHBP3 expression began to be up-regulated at the 4th instar larva stage and up-regulated to 3.87 fold.Ms JHBP5 was highly expressed at the 3rd instar larva stage,and then down-regulated,and the pre-pupal stage began to be regulated,and the expression reached 12.65 fold in the pupal stage.Ms JHBP7 also had high expression at the 4th instar larva stage and reached the highest expression of 10.56 fold at the pre-pupal stage.(4)After interfering Ms JHBP3,the content of juvenile hormone was 1.14 fold,and the molting hormone rate and pupation rate were significantly reduced.The emergence rate decreasing by 7.42%and the pupation rate decreasing by 3.75%.After interferencing Ms JHBP5,the content of juvenile hormone was 1.06 fold,and the molting hormone rate was not significant.The pupation rate decreased significantly by 1.63%.the contents of juvenile hormone and molting hormone were determined,and the juvenile hormone content was adjusted upwards and the content of molting hormone was down.After interfering with Ms JHBP3 and Ms JHBP7,the weight of M.separata increased,but there was no significant difference compared with the control.(5)After transcriptome analysis,a total of 52.280 Gb of Clean Data was obtained,the average amount of Clean Data for each sample was 8.713 Gb.A total of 212 differential c DNA sequences were obtained,including 105 up-regulated sequences and 107 down-regulated sequences.Among them,the juvenile hormone acid methyltransferase,juvenile hormone esterase,juvenile hormone epoxide hydrolase were all up-regulated.The differential c DNA sequences were mostly related c DNA sequence that had an impact on growth and development,including JH synthesis c DNA sequence and JH degradation c DNA sequence,which affected multiple growth and development pathway c DNA sequence after interferencing.The enrichment results also showed that it was closely related to growth and development.Studies have shown that Ms JHBP3 affects the growth and development of M.separata and is involved in regulating JH metabolism.The results of this experiment provide basic data for mining key c DNA sequence of M.separata growth and development,and provide new target c DNA sequence and approaches for M.separata biological control.