The Function Of Cytochrome P450 Reductase And Cytochrome P450 Genes In Locusta Migratoria And Ostrinia Furnacalis

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Transcriptome data assembly,gene function annotation and functional analysis of cytochrome P450 gene in the Asian corn borerCytochrome P450 monooxygenases(P450s)are a large superfamily,catalyzing at least 60 types of chemically distinct reactions.Insect P450 s plays a key role in degradation of insecticides,and biotransformation of plant toxics.P450 have been well studied in model insects,but less is known about P450 family in Ostrinia furnacalis.In order to study the effect of CYP450 in Ostrinia furnacalis,SRA method was used to analyze the 8 groups of transcriptome,then turn to Biomarker.The 36.54 Gb clean data were obtained by Trinity software.The percentage of Q30 base of each sample was not less than 87.24%,and the mean value of GC% was 49.74%.The transcriptome sequencing data saturation test showed that the number of genes detected was becoming saturated.The total 59911 unigenes were obtained after assembly and the total length of the sequence is 44249132 bp,among them 24 517unigenes(38.33%)were annotated in the public databases Nr,Swiss-Prot,COG,KOG,Pfam,GO and KEGG.The average length was 738.58 bp and N50 was 1239 bp.31P450 full-length genes were obtained,and the classification and naming of gene families were carried out.Six P450 genes were selected from the obtained full-length sequence,and the function was studied by using RNAi technology.??Cloning and expression profiling of Cytochrome P450 Reductase Gene from the Asian Corn Borer,Ostrinia furnacalisCytochrome P450 reductase(CPR)is a redox partner of cytochrome P450,and plays important roles in cytochrome P450-mediated metabolism of endogenous and exogenous compounds.In this study,the full-length cDNA(2323 bp)of a CPR gene was cloned from Asian corn borer,Ostrinia furnacalis and named OfCPR(GenBank accession No.: MG255127).It contained a 2049 bp open reading frame,encoding a putative protein of 682 amino acids with a predicted molecular weight of 76.9 kDa and the theoretical pI of 5.57,respectively.The result of homology modelling showed that OfCPR was consisted of one membrane anchor region,and three conserved domains(FMN binding domain,connecting domain,and FAD/NADP bindingdomain).Phylogenetic analysis showed that OfCPR was grouped in the Lepidoptera branch and was closely related to the CPRs from Diptera and the Optera insects.The OfCPR gene was ubiquitously expressed at all tested-developmental stages and was the most abundant in the adult stage,and higher expressed in first-and fifth-larval stages.The OfCPR gene was expressed on each day of the fifth-larval stage,and was the highest in the third day of fifth-larval stage.Tissue-specific expression analysis showed that OfCPR gene was the highest expressed in the gut.This manuscript laid a foundation for elucidating the function of the OfCPR gene,and will be conducive to prevention and control of Ostrinia furnacalis.? ? Knockdown of NADPH-cytochrome P450 reductase increases the susceptibility to carbaryl in the migratory locust,Locusta migratoriaHowever,the CPR from Locusta migratoria has not been well characterized and its function is still unclear.The full-length of CPR gene from Locusta migratoria(LmCPR)was cloned by RT-PCR based on transcriptome information.The catalytic residues,FAD binding motif,membrane anchor domain,and 3 conserved binding domains(FMN,FAD,and NADP)were analyzed by bioinformatics analysis and homology modeling.Phylogenetic analysis showed that LmCPR was grouped in the Orthoptera branch and was more closely related to the CPRs from hemimetabolous insects.The LmCPR gene was ubiquitously expressed at all developmental stages and was the most abundant in the fourth-instar nymphs and the lowest in the egg stage.Tissue-specific expression analysis showed that LmCPR was higher expressed in the antenna,ovary,hindgut,and integument.Silencing of LmCPR obviously reduced the enzymatic activity of LmCPR,and enhanced the susceptibility of Locusta migratoria to carbaryl.Conclusion: These results suggest that LmCPR contributes to the susceptibility of L.migratoria to carbaryl and could be considered as a novel target for pest control.??Induction of LmP450 by plant allelochemicals and detoxification mechanismBased on the preliminary study of the research group,we got five CYP450 genes induced by xanthotoxin.RNAi and leaf-dipping methods were carried out to show that Lm6FE1 and Lm6FD1 defensed against xanthotoxin.Then we develop the research on the induction mechanism.Firstly,whether the P450 genes,which weresignificantly induced by multiple concentration of xanthotoxin,were regulated by transcription factors under physiological conditions or not? Secondly,how about the response of P450 s induced by xanthotoxin.Conclusion: CYP6FE1 and CYP6FD1 are regulated by CncC;CYP6FD1 are regulated by Maf.In addition,silencing of CncC enhanced the susceptibility of Locusta migratoria to xanthotoxin;Silencing of Maf reduced the susceptibility of Locusta migratoria to xanthotoxin.These dates showed that Locusta migratoria emplys metabolic detoxication through transcription factors CncC and Maf to regulate multiple P450 genes and detoxify xanthotoxin.