The Cloning And Functional Analysis Of AOX、ATF And DES Genes Of Apolygus Lucorum

Apolygus lucorum is one of the key species of mirid pests on cotton, which currently mainly rely on chemical control. Studying functions of important genes of miridae is significant to explore the green and safe control measures. Sex pheromones of miridae are mainly comprised of short carbon chain of aldehydes and esters. Previous studies have reported alcohol oxidase(alcohol oxidase, AOX), acetyltransferase(acetyltransferase, ATF) and desaturase(desaturase, DES) is an important enzymes of the synthetic process of aldehydes and esters. But their function in Apolygus lucorum not clear, so this paper carried out the function of these three genes in green bugs in Apolygus lucorum.RNAi injection interference is an important means of studying gene functions in this paper. We firstly studied the optimum injection volume of RNAi, the result shows that the optimum injection volume of the first day of emergence of adult Apolygus lucorum is 100 n L.The paper studied 8 ATF genes, 2 AOX genes and 1 DES gene’s expression differences in the scent glands of sex pheromone release peak male and female and immature male and female. We found a specific gene NAA25 of ATF in 3 instar Lygus scent glands of female specific high expression by fluorescence quantitative PCR. This gene may be associated with the synthetic sex pheromone of Apolygus lucorum.We interfere NAA25、NAA20 and NAA15 genes by microinjection. After the silence of NAA25, the number of mature follicles of sexually mature female Apolygus lucorum(Meyer-Dür.) reduced by 87.1%(p <0.005). After the silence of NAA20, the number of mature follicles of sexually mature female Apolygus lucorum(Meyer-Dür.) reduced by 95.6%(p <0.005). After the silence of NAA15, the number of mature follicles of sexually mature female Apolygus lucorum(Meyer-Dür.) reduced by 98.8%(p <0.005). We also use real-time quantitative PCR to detect the expression levels of these three genes in whole insects from the first day of emergence to the day of sexual maturity, these three genes are highly expressed at the first day of emergence and the day before sexual maturity. Therefore, we speculate these three genes play an important role in the ovarian development.In the study, we obtained 2 full-length N-alpha-acetyltransferase(NAA) by RACE, of which NAA25 belong to Nat B enzyme complex of two subunits, NAA15 subunit belongs to Nat A complex enzyme. NAA25 has a full-length sequence of 2898 bp, of which ORF region is 2064 bp, encoding 687 amino acids. NAA15 has a full-length sequence of1103 bp, of which ORF region is 759 bp, encoding 252 amino acids.