Discovery Of Eco-friendly Insecticide Targets Based On The Enzymes In Juvenile Hormone Biosynthesis

Chemical insecticides have made a major contribution to agriculture and health.However,as a result of improper use and the inherent shortcomings of some insecticides,many insecticides showed negative impacts on the ecological environment.Therefore,eco-friendly insecticides have received global attention in recent years.RNA interference is a method which can be used in gene functional identification.Recent research indicated that using RNAi to discover key gene in the development and reproduction of pests can be an effective way to find new insecticide targets.Juvenile hormone(JH)is an essential hormone in insect development.The enzymes involved in JH biosynthetic pathway have the potential to be insecticide targets.In our research,11 related JH synthesitic enzymes of Manduca sexta were selected as the target genes.The key genes in this pathway were discovered based on the results of tissue distribution and developmental expression.Sequence alignment was performed to determine potential enzymes or receptors which can be eco-friendly pesticide targets.FOLD,JHAMT and CYP15C1 appeared to be eco-friendly insecticide target candidates.The corpora allata of M.sexta larvae was incubated with FOLD,JHAMT and CYP15C1 dsRNAs,respectively.Their expressions were down by 45.5%,93.8%and 64.5%,respectively.Their JH biosynthesis was obviously decreased by 64.4%(FOLD),70.6%(JHAMT),75.4%(CYP15C1).Thus,FOLD,JHAMT and CYP15C1 can be the potential eco-friendly insectide targets due to their important role in insect development and their specificity to other species.Greanylgeranio(FOLD inhibitor),Sinefugin(JHAMT inhibitor)and Spinosyn(CYP15C1 inhibitor)show significant JH biosynthesis inhibitions(their inhibition at 10 ?M was 62.1%,58.5% and 63.5%,respectively.These inhibitors have the potential to be lead compounds for new insecticide discovery.Our study has identified the key gene in JH biosynthesis,verifed the validity and safety of the target candidates.FOLD,JHAMT and CYP15C1 were found as targets for insecticide design.The enzyme inhibitors have been tested to discover new insecticide lead compounds.Furthermore,the methods we used in our study can provide an empirical strategy for eco-friendly insecticides discovery.